Polyhydrophenanthrene diols

ABSTRACT

Alkyl polyhydro-2-phenanthrylideneacetates are prepared by interacting the corresponding 2-oxopolyhydrophenanthrenes with a tri-lower-alkyl Alpha -phosphono-lower-alkanoate. Said alkyl polyhydro-2-phenanthrylideneacetates are hydrolyzed to the free acid, and then reesterified via the acid chloride with a tertiary-amino-lower-alkanol to give basic esters having cardiotonic activity.

United States Patent Shaw et al. 1 Nov. 25, 1975 [5 1 POLYHYDROPHENANTHRENE DIOLS 2,750,425 6/1956 Subleskey 260/617.5 3,592,838 7/1971 Shaw et al. 260/617.5

Inventors: Philip E. Shaw, Winter Haven, Fla.;

S01 J. Daum, Albany; Robert L. Clarke, Bethlehem, both of N.Y.

Assignee: Sterling Drug Inc., New York, N.Y.

Filed: Aug. 31, 1973 Appl. No; 393,623

Related U.S. Application Data Division of Scr. No. 269,402, July 6, 1972, Pat. No. 3,830,843, which is a division of Ser. No 879,920, Nov. 25, 1969, Pat. No. 3,755,361, which is a continuation-in-part of Ser. No. 585,762, Oct. 11, 1966, Pat. No 3,592,838.

[52] U.S. Cl 260/617.5; 195/104; 260/456 R; 260/456 P; 260/590; 260/611 A; 424/246; 424/274; 424/277; 424/278; 424/305; 424/308; 424/317; 424/320; 424/325 [51] Int. Cl. C07C 35/42 [58] Field of Search 260/617.5

[56] References Cited UNITED STATES PATENTS 2,722,532 11/1955 Arth et al. 260/6175 FOREIGN PATENTS OR APPLlCATlONS 763,208 12/1956 United Kingdom 260/617 Primary Examiner.loseph E. Evans Attorney, Agent, or FirmThomas L. Johnson; B. Woodrow Wyatt [57] ABSTRACT Alkyl polyhydr0-2-phenanthrylideneacetates are prepared by interacting the corresponding 2- ox opolyhydrophenanthrenes with a tri-lower-alkyl a-phosphono-lower-alkanoate. Said alkyl polyhydro-2- phenanthrylideneacetates are hydrolyzed to the free acid, and then reesterified via the acid chloride with a tertiary-amino-lower-alkanol to give basic esters having cardiotonic activity.

3 Claims, No Drawings POLYHYDROPHENANTHRENE DIOLS This application is a division of our copending application Ser. No. 269,402, filed July 6, 1972, now U.S. Pat. No. 3,830,843, which is in turn a division of our copending application Ser. No. 879,920, filed Nov. 25, 1969, now U.S. Pat. No. 3,755,361, which is in turn a continuation-in-part of our prior copending application Ser. No. 585,762, filed Oct. 11, 1966, now U.S. Pat. No. 3,592,838.

This invention relates to tricyclic substituted acids and esters thereof, and in particular is concerned with polyhydro-2-phenanthrylideneacetic acids and basic esters thereof, and with intermediates in the preparation thereof.

One aspect of the invention is concerned with compounds of the formula wherein:

R is hydrogen, lower-alkyl or amino-lower-alkyl;

R is hydrogen, lower-alkyl or hydroxy;

R and R are hydrogen or lower-alkyl;

X is H (H)(OI-I), (H)(O-acyl) or and Z is O=C, (l-IO)CI-I, (acyl-O)CI-I, CH (lower-alkyl)- (lower-alkoxy)CH, (halogen)CH, H NC(=NH)NI-IN=C, (O NO)Cl-I, (C l-I CH S) C, (C I-I CI-I SO C, [(lower-alkyl);, N]CI-1, (piperidino)CI-I, (pyrrolidino)CH, (4-hydroxypiperidino)CI-I,

where n is 2 or 3. Also included are compounds of formula I having a double bond in the 8,8a-position or the 8a,9-position; compounds of formula I wherein the exocyclic double bond is saturated; and compounds of formula I having two identical lower-alkyl groups in the 8-position.

The term lower-alkyl used above in defining the groups R, R" and R and certain subgroups under Z stands for alkyl groups having up to about six carbon atoms, including such groups as methyl, ethyl, propyl, isopropyl, butyl, hexyl, and the like.

When R in formula I stands for an amino-lower-alkyl group, it represents a lower-alkyl group substituted by a basic amino group. The exact nature of the basic amino moiety is not critical, although it is preferred that it have a molecular weight less than about 200. A particularly preferred type of amino-lower-alkyl group has the structure -Y-N=B wherein Y is lower-alkylene of 2-5 carbon atoms and N=B is unsubstituted amino, lower-alkylamino, di-lower-alkylamino, polymethylenimino of 5-7 ring carbons and lower-alkylated 2 derivatives thereof, 4-morpholinyl and lower-alkylated derivatives thereof, or l-piperazinyl and loweralkylated derivatives thereof. The term lower-alkyl as used in defining portions of the amino moiety N=B stands for alkyl groups having up to about six carbon atoms.

When Z in formula I above stands for (acyl-O )CI-I, or X stands for (H)(O-acyl), the acyl groups are carboxylic acyl groups having from one to twelve carbon atoms and molecular weights less than about 250. Representative of the lower-carboxylic acyl radicals which can be present are lower-alkanoyl, e.g., acetyl, propionyl, isobutyryl, caproyl, heptanoyl, octanoyl, dodecanoyl, trimethylacetyl, and the like; cycloalkyl-loweralkanoyl wherein cycloalkyl has 5-6 ring members, e.g., B-cyclopentylpropionyl, B-cyclohexylpropionyl, and the like; benzoyl; phenyl-lower-alkanoyl or -alkenoyl, e.g., phenylacetyl, B-phenylpropionyl, cinnamoyl, and the like; phenoxy-lower-alkanoyl, e.g., p-chlorophenoxyacetyl; carbamyl, including unsubstituted carbamyl, N-lower-alkylcarbamyl, N-phenylcarbamyl and N,N-di-lower-alkylcarbamyl; and pyridylcarbonyl, e.g., nicotinoyl and isonicotinoyl. In acyl radicals containing a phenyl group, the benzene ring thereof can be unsubstituted or substituted by any number and kind of substituents inert under the reaction conditions used, including lower-alkyl, for example p-tolyl; lower-alkoxy, for example 3,4-dimethoxyphenyl; halogen (including fluorine, chlorine, bromine and iodine), for example 2-bromophenyl; and nitro, for example p-nitrophenyl. The lower-alkyl and lower-alkoxy groups preferably have from one to four carbon atoms.

The invention is not limited to any particular stereochemical configuration of the compounds of formula I, although a preferred configuration of the ring system is trans-anti-trans (4aoz,4b,8,8aa,l0aB), such compounds being derived from readily available starting materials.

A further aspect of the invention is concerned with compounds of the formula wherein:

R is hydrogen, lower-alkyl or amino-lower-alkyl;

R is hydrogen or lower-alkyl; and

R' is hydrogen, lower-alkyl or carboxylic acyl of 1-12 carbon atoms.

The terms lower-alkyl, amino-lower-alkyl and acyl have the meanings given above. The invention is not limited to any particular stereochemical configuration of the compounds of formula II, although a preferred configuration is 4aa,10aB, such compounds being derived from readily available starting materials.

The compounds of formulas I and I1 wherein R is lower-alkyl are prepared from compounds of formulas III and IV, respectively, including compounds of formula 3 III having a double bond in the 8,8a-position, Z being C=O, and compounds of formula III having two identical lower-alkyl groups in the 8-position and a double bond in the 8a,9-position, Z being C=Oz III RIIIO by reacting compounds of formulas III and IV with a tri-lower-alkyl a-phosphono-lower-alkanoate of the formula in the presence of a basic compound capable of acting as a proton acceptor. Examples of such basis compounds are alkali metal alkoxides, amides or hydrides, e.g., sodiumethoxide, sodium amide or sodium hydride, and the reaction is preferably carried out in nonaqueous medium at room temperature. The compounds of formula I where X is O can be prepared in this manner because the lO-oxo group is relatively unreactive compared to the 2'-oxo group.

It is preferred to employ compounds of formula III wherein Z is (HO)CI-I, (acyl-O)CH, CH (loweralkyncunfi (lower'alkoxy)cH (halogemCH of the compounds where R is hydrogen with dehy- (C6H5CH2S)2C" 45 droabietylamine.

The intermediates of formula IV can be prepared as" (CH C (CH C illu trated 'n the follo n flo sheet 2.. r 2n S s l w1 g w O-lowereallcyl 0 'CH CH i (011 M c 2 c=o BI'CH CHQ base m 0/ (JH Cl-i COO1ower-allryl O CH 'CH (CH c 2 c=o ii so 0 cH -c-coo-1ower-a1ic 1 CH CK 4 in the reaction with a tri-lower-alkyl a-phosphono-lower-alkanoate; however, Z can also be O=C if a double bond is present at the 8,8a-position; or two identical f lower-alkyl groups are in. the 8-position. Thecompounds of formula I wherein Z is O=C, (acyl-0)CH,-

[(lower H NC(=NH)NHN=C, (O NO)CH, alkyl) N]CI-I, (piperidino)CI-I, (4-hydroxypiperidino)CH, or .(pyrrolidino)CI-I can readily be prepared from the compounds of formula I wherein Z is i (HO)CH or O=C by conventional means as illustrated in the examples below. I

The reaction of compounds of formulas III andIV with a tri-lower-alkyl a-phosphono-lower-alkanoate gives a mixture of geometric isomers (cis and trans) involving the groups about the exocyclic double bond. Although in many instances it is possibleto separate. the isomers by physical means, it is not essential to the present invention that the isomers be separated .be-

cause there is little difference in the physiological properties of the isomers.

The compounds of formulas I and II wherein R is hydrogen are prepared by alkaline hydrolysis of the com.- 2 pounds of formulas I and II wherein R is lower-alkyl. I

The compounds of formulas I and II wherein R is amino-lower-alkyl are prepared by esterification of the compounds of formulas I and 11 wherein R is hydrogen by employing the appropriate amino-lower-alkanol or amino-lower-alkyl halide. A preferred method comprises reacting the acid halide ofa free acid of formulas I or II with a tertiaryamino-lower-alkanol, although an alternative procedure involves reacting an alkali metal salt (e.g., sodium salt) of the acid with an amino-.loweralkyl halide (e.g., chloride or bromide). Surprisingly, the acid halide process can be used even in the presence of a free hydroxy group [Z=(HO)CI-I It appears that if reaction does occur at the hydroxy group during acid halide formation, the hydroxy group is regenerated during the preparation of the basic ester.

The products of formulas I and II are obtainedin the form of racemic mixtures of optically activedand 1 forms. If desired, the latter can be separated by conventional resolution procedures, for example, by treatment I 0-10wer-alkyl A Z-carbo-lower-alkoxycyclohexane-1,4-dione 4- ketal (m 2 or 3) is aralkylated with a m-lowersalkoxyphenylethyl bromide, and the resulting product V is cyclized with acid to yield 3,4,9,lO-tetrahydro-7-loweralkoxy-2(II-l)-phenanthrone (VI). Catalytic hydrogenation of VI with palladium-on-carbon gives a compound of formula lV having the 4aoz,lOaoz-configuration (IVA). Sodium borohydride reduction of VI gives 1,2,3,4,9,10-hexahydro-7-methoxy-2-phenanthrol (x ll). Sodium-liquid ammonia reduction of the latter provides a mixture of 1,2,3,4,4aa,9,10,10aB-octahydro-7-lower-alkoxy-2fiand Zoz-phenanthrol (VIIIC and VIIID), which can be oxidized with chromic oxide to give a compound of formula IV having the 4aa,l- @aB-configuration (IVB). Sodium borohydride reduction of IVA gives a mixture of 1,2,3,4,4az,9,10,10aaoctahydro-7-lower-all oxy-2B- and 2a-phenanthrol (VIIIA and VIIIB).

The compounds of formula III wherein R and R are hydrogen, X is H and Z is (HO)Cl-l are prepared by reduction of the octahydrophenanthrenes of formulas VllIA-D, as follows:

l) Iii/NH 9 2) Hydrolysis VIII A compound of formula VIII is first reduced with lithium and liquid ammonia in the presence of alcohol (Birch reduction) followed by hydrolysis to give a 4-,4a,4b,5.6,7,8,8a,9,10-decahydro-7-hydroxy-2(3H)- phenanthrone (IX), which can then be reduced further either catalytically or by alkali metal-ammonia reduction to give 3,4,4a,4b,5,6,7,8,8a,9,l0,10a-dodecahydro-7-hydroxy-2(1H)-phenanthrone (X) (III; Z is (HOMIH, R and R" H, X H, The stereochemistry of the ring system and the 7-hydroxy group is dependent upon the particular VIII isomer used and the nature of the reduction of IX, as will be apparent from the specific examples below. The function groups of X can also be altered by conventional means as will b: i1 lustrated by the examples below.

The compounds of formula III wherein R is methyl can be prepared from the compound:

XI (R' by reductive alkylation with lithium in liquid ammonia and a lower-alkyl halide.

The compounds of formula III wherein R. is OH or X is O can be prepared by microbiological oxidation of compound IX above. When IX is incubated with acuiture medium of an organism of the genus Cunninghamella there is obtained a mixture of the hydroxylated products: I

and

XII

XIII

Treatment of XII with sulfuric acid in acetic acid converts it by rearrangement to a compound of formula III 1 I where Z is (CH COO)CH, R is H, R" is H and Xis. =0. Oxidation of XIII with chromic oxide converts it to a compound of formula III wherein Z is O=C, R is OH,

R is H, X is H and there is a double bond in the 8,8a-

position.

The compounds of formula III wherein there are two lower-alkyl groups in the 8-position and a double bond in the 8a,9-position, Z being O=C, are prepared as follows:

lower-allcyl iower-alIcyl XIV lower-allql "lower-allcyl XVI Alkylation of XIV (R is hydrogen or lower-alkyl) with a lower-alkyl halide in the presence of potassium tertiary-butoxide gives a hydroxy ketone of formula XV. The latter can be oxidized with chromic oxide to the diketone XVI. The IO-oxo group can then be introduced by the action of tertiary-butyl chromate to give the triketone XVII (III; R is H or lower-alkyl, R" is H, X is 0, Z is O=C, 8,8-di-lower-alkyl, A Alternatively, the compounds XV or XVI can be oxidized with tertiary-butyl peracetate in the presence of cuprous chloride to introduce an acyloxy group into the position. Hydrolysis and oxidation of the latter then gives the triketone XVII.

Further starting materials for the preparation of compounds of formula I having two identical lower-alkyl groups are of the formula lower-allq'l "lcwer-allcyl XVIII where R is hydrogen or lower-alkyl. The compounds of formula XVIII can be prepared from compounds of formula XVI by forming the mono-ethylene glycol ketal of the 2-oxo group, reducing the 8-oxo group with lithium aluminum hydride to an SB-hydroxy group, reducing the 8a,9-double bond with hydrogen in the presence of palladium-on-carbon, and finally cleaving the 2-ketal group.

The compounds of formula I where the exocyclic double bond is saturated can be prepared by hydrogenation (palladium-on-carbon) of the unsaturated analogs where R is lower-alkyl.

The structures of the compounds of the invention were established by the modes of synthesis, by elementary analysis, by interpretation of their infrared, ultraviolet and NMR spectra, and by their behavior in thin layer chromatography (TLC) and gas-liquid phase chromatography (glpc).

lower-alkyl iower-alkyl XVII The compounds of formulas I and II wherein R is amino-lower-alkyl are basic in nature and form acidaddition salts with moderate to strong inorganic or organic acids. For pharmacological purposes it is preferred to use water-soluble, pharmaceutically acceptable acid-addition salts, although all acid-addition salts are useful as characterizing derivatives of and as intermediates in the purification of the free bases.

Pharmacological evaluation of the basic ester compounds of the invention has shown that they possess cardiotonic activity similar to that shown by the alkaloid cassaine and are thus useful in increasing the ventricular contractile force of the mammalian heart. The compounds are prepared for use in the form of a sterile aqueous solution of a water-soluble, pharmaceutically acceptable acid-addition salt. The amidinohydrazone derivatives of the intermediate tricyclic ketones also have similar cardiotonic activity.

Chemotherapeutic evaluation has shown that the compounds of formula I wherein R is hydrogen possess bacteriostatic activity.

The following examples will further illustrate the invention without the latter being limited thereby.

EXAMPLE 1 2-Carbomethoxy-2-(m-methoxyphenethyl)-cyclohexane-l,4-dione 4-ethylene ketal [V; R' is CH A solution of 5.33 g. (0.047 mole) of potassium tbutoxide in ml. of t-butanol was stirred while 10.0 g. (0.047 mole) of 2-carbomethoxycyclohexane-l,4- dione 4-ethylene ketal in, ml. of t-butanol was added. An additional 100 ml. of t-butanol was added, followed by 10 g. (0.047 mole) of m-methoxyphenethyl bromide. The suspension was refluxed and stirred mechanically for 72 hours. At this point the reaction mixture was slightly alkaline to pH paper. The reaction mixture was cooled and added to ice water. Ether was added followed by dilute hydrochloric acid. The layers were separated, and the ether layer was washed with a saturated sodium bicarbonate solution. The ether was extracted twice with 200-ml. portions of 5 percent aqueous potassium hydroxide. The aqueous alkaline solution on acidification with hydrochloric acid yielded 1.45 g. of starting 2-carbomethoxycyclohexane-1,4- dione 4-ethylene ketal which was identified by its melting point. The ether solution from which the starting material had been removed was then washed with ice cold dilute hydrochloric acid, saturated sodium bicarbonate solution and saturated sodium chloride. After drying the ether solution (Na SO the solvent was removed to leave a yellow viscous oil. This residue was distilled and the fraction that boiled at l90205C. (0.08-0.15 mm.) was collected to give 10.1 g. of 2-carbomethoxy-2-(m-methoxyphenethyl)-cyclohexane- 1,4-dione 4-ethylene ketal. A redistilled sample boiled at 195-196C. (0.18-0.20 mm.); n,,. 1.5267.

EXAMPLE 2 3,4,9,10-Tetrahydro-7-methoxy-2(1H)-phenanthrone 2; R is CH Sulfuric acid (1 liter 1 IN) was added all at once to a solution containing 115.4 g. (0.33 mole) of 2-carbometl10xy-2-(m-methoxyphenethyl )-cyclohexane- 1 ,4- dione 4-ethylene ketal (Example 1) in 1 liter of dioxane. The reaction mixture was stirred at 100C. in a nitrogen atmosphere for four and one-half hours. The cooled solution was poured into ice and enough water was added to bring the total volume to 3 liters. The aqueous mixture was extracted with ether several times. The combined ether layers were washed with 10 percent sodium carbonate and finally with saturated sodium chloride solution. The ether solution was dried (Na SO and evaporated by warming in vacuo to afford 73.9 g. of a red oil. This material was chromatographed on 2 kg. of silica gel. Elution with 4 liters of ether-pentane (1:9) and 20 liters of ether-pentane (1:3) removed less polar material. Continued elution with 5.5 liters of ether-pentane (1:3) afforded 18.2 g. of an oil which crystallized from ether containing hexane to give 16.5 g. of 3,4,9,l-tetrahydro-7-rnethoxy- 2(ll-l)-phenanthrone, m.p. 7577C. Further elution (5 liters) gave an additional 17.8 g. of the compound.

EXAMPLE 3 1,2,3,4,9, l 0-Hexahydr0-7-methoxy-2-phenanthrol [\lll; R' is CH A solution of 36.7 g. (0.16 mole) of 3,4,9,lO-tetrahydro-7-methoxy-2(1H)-phenanthrone (Example 2) in 1500 ml. of 95 percent ethanol was treated with 19 g. of sodium borohydride in 150 ml. of water. After three hours at room temperature, acetone was added to the reaction mixture to decompose the excess borohydride. The volume was reduced to about 400 ml. by warming in vacuo. Water and ether were added and the layers were separated. The ether was washed with saturated sodium chloride solution. The aqueous layers were washed again with fresh ether and the organic layers were then combined. The ether solution was dried (Na SO.,) and the solvent was removed leaving behind an oily residue (36.1 g.). Crystallization from ether containing hexane gave 20.3 g. of 1,2,3,4,9,10-hexahydro-7-methoxy-2-phenanthrol, m.p. 9394C. Further concentration of the mother liquor afforded another 9.3 g. of the compound, m.p. 90-92C.

EXAMPLE 4 3,4,4aa,9,10,10aot-Hexahydro-7-methoxy-2(11l)-phenanthrone [1VA; R is CH A solution of 1 g. (4.4 mmoles) of 3,4,9,10-tetrahydro-7-methoxy-2(1H)-phenanthrone (Example 2) in 300 ml. of ethyl acetate was hydrogenated at a pressure of 3.86 kg./cm and room temperature in the presence of 100 mg. of 10 percent palladium-on-carbon. After 1 hour the hydrogenation was complete and the catalyst was removed by filtration. The solvent was removed by warming in vacuo and the residue was crystallized from ether containing hexane 3,4,4a0:,9,10,l0aa-hexahydro-7-rnethoxy-2(1H).-phenanthrone, m.p. 92-93C. and a second crop mg), m.p. 8991C.

EXAMPLE 5 I 1,2,3 ,4,4aoz,9,1 0, l 0aac-Octahydro-7-methoxy-2B-' phenanthrol and -2a-phenanthro1 [VlllA and B; R is 7 (Method 1) A solution of 15 g. (0.066 mole) of 1,2,3 ,4,9,10-hexahyclro-7-methoxy-2-phenantl1r01 (Example 3) in 300 ml. of ethyl acetate was hydrogenated at a pressure of 3.86 kg/em and room temperature in the presence of 1.5 g. of 10 percent palladium-on-carbon. After 45 minutes the hydrogenation was complete and the catalyst was removed by filtration. The filtrate was combined with the filtrate of an identical experiment. The solvent was removed by warming in vacuo to leave 31.5 g. of an oily residue which crystallized from ether containing hexane. Recrystallization of this material from ether containing hexane afforded 13.6 g. of

solid, m.p. 80-83C.'

The mother liquor was chromatographed on 1.5 kg. of silica gel. Elution with 13 liters of ether-pentane (l :1) removed some less polar material. Continued elution with 5.5 liters of solvent of the same concentration.

afforded 6 g. of material which, on crystallization from ether containing hexane, yielded 4.83 g. of solid, m.p.

8487C. and 0.4 g. of solid,m.p. 8185C. Combina- I tion with the original solid obtained by direct crystallization and recrystallization from ether containing hexane afforded 16.1 g. of 1,2,3,4,4aoz,9,10,lOaa-octahydro-7-methoxy-2,8-phenanthrol (VIIIA), m.p. 8488C. This compound apparently shows polymorphism. Recrystallization from ether containing hexane gave material melting at 7782C. 7

Further elution in the above described chromatography, with 2.5 liters of ether-pentane (1:1 gave 4.2 g. of a mixture of two compounds. Finally, elution with another 4 liters of the same solvent mixture gave an oily residue which crystallized from ether containing hexane and afforded 5.3 g. of 1,2,3,4,4a'a,9,l0,10aaoctahydro-7-methoxy-2a-phenanthrol (VIIIB),

ane gave 5 g., m.p. 8990.5C.

(Method 2) A solution of 580 mg. (2.5 mmoles) of 3 ,4,4aa,9, l 0, l Oaa-hexahydro-7-methoxy-2( 1H )-phenanthrone (Example 4) in 25 ml. of absolute ethanol was treated with 240 mg. of sodium borohydride in 2.5 ml. of water. The reaction mixture was kept at room coated plates which were developed with ether-pentane (3:1). About mg. of residue was put on each of 3 plates (20 cm X 40 cm) carrying a l-mm. coating of silica gel. The major band from the plates (less polar) afforded 310 mg. (54 percent) 1,2,3 ,4,4aa,9, 1 0, l Oaa-octahydro-7-methoxy-2B phenanthrol (VlllA), which was identified by melting point, infrared spectrum and TLC analysis. The more polar band afforded 1 10 mg. 19 percent) of l,2,3,4,4aa,9,10,lOaa-octahydro-7-methoxy-2am.p. 6770C. Recrystallization from ether containing hex- 13 phenanthrol (VIllB) which was identified by melting point, infrared spectrum and TLC analysis.

EXAMPLE 6 1,2,3 ,4,4aa,9,l0,10aB-Octahydro-7-methoxy-2B- phenanthrol and -2a-phenanthrol [VlIIC and D; R' is CH Sodium (3.15 g., 0.137 mole) was dissolved in 1 liter of liquid ammonia. A solution of 15 g. (0.066 mole) of 1,2,3,4,9,10-hexahydro-7-methoxy-2-phenanthrol (Example 3) in 100 ml. of tetrahydrofuran and 25 ml. of aniline was added over a twenty minute period to the ammonia solution which was being stirred. At the end of the addition 14 g. (0.26 mole) of ammonium chloride was added portion-wise during minutes to discharge the blue color. The ammonia was evaporated and water and ether were added to the residue. The aqueous layer was separated and extracted twice with ether. The combined ether layers were washed three times with water, twice with 3N hydrochloric acid, three times with saturated sodium chloride solution, dried (Na SO and the solvent was removed to leave an amber colored residue. After crystallization and several recrystallizations from ether containing hexane, 4.7 g. of 1,2,3,4,4aa,9,10,10aB-octahydro-7-methoxy- Zap-phenanthrol (VIIIC), m.p. 9899C. was obtained.

The combined mother liquors were chromatographed on 5 kg. of silica gel. Preliminary elution with ether-pentane (1:1) separated some less polar oils which were discarded. Continued elution with etherpentane (1.1:1) afforded first g. of crude 1,2,3 ,4,4aa,9,10,10aB-octahydro-7-methoxy-2B- phenanthrol. Continued elution with the same solvent system next afforded 12 g. of crude 1,2,3,4,4aoz,9,10,- lOaa-octahydro-7-methoxy2/3-phenanthrol (VlllA). Further elution with ether-pentane (1.5:1 finally gave 53 g. of an oil which yielded 13 g. more of 1,2,3,4,4aa,9,10,10aB-octahydro-7-methoxy-2aphenanthrol (VlllD), m.p. 98l00C.

Recrystallization of the crude 1,2,3,4,4aa,9,10,l0aB- octahydro-7-methoxy-2B-phenanthrol from ether afforded 13.75 g. of this product.

A similar chromatographic separation afforded 1 ,2,3,4,9,10-hexahydro-7-hydroxy-2-phenanthrol [V1]; R is H], m.p. 196.5l98C. (10 percent) when 100 percent ether was put through the column.

EXAMPLE 7 3,4,4aa,9, l 0, lOaB-Hexahydro-7-methoxy-2( 1H )-phenanthrone [IVB; R' is CH A solution of 25 g. 0.11 mole) of l,2,3,4,4aa,9,10,- 10aB-octahydro-7-methoxy-2a-phenanthrol in 500 ml. of pyridine was added to a suspension of 25 g. of chromium trioxide (0.25 mole) in 500 ml. of pyridine. After standing at room temperature for 68 hours, the reaction mixture was added to 1.5 liters of ethyl acetate. The chromium salts were removed by passing the suspension through infusorial earth. The solvent was removed by heating in vacuo. Ether was added to the viscous residue and the mixture was filtered again. Evaporation of the ether afforded g. of viscous oil. Crystallization from cyclohexane containing ether afforded 6 g. of 3,4,4aa,9,10,10aB-hexahydro-7-methoxy-2(1H)- phenanthrone, m.p. 62-63C. Concentration of the mother liquor afforded another 4 g., 5962C. Chromatography of the mother liquors on silica gel and elution with ether-pentane (1:3) afforded an additional 6.14 g. of 3,4,4aa,9,l0,10aB-hexahydro-7-methoxy- 2(1H)-phenanthrone, m.p. 64.566C. when recrystallized from ether-hexane.

EXAMPLE 8 4,4aa,4bB,5,6,7,8,8aB,9, lO-Decahydro-7a-hydroxy- 2(3H)-phenanthrone [lX; 7(1,4aa,4bB,8aB-configuration] A solution of 11 g. (0.05 mole) of l,2,3,4,4aa,9,10,- l0aa-octahydro-7-methoxy-2B-phenanthrol (VIllA,

Example 5) in 200 ml. of tetrahydrofuran and 200 ml.

of t-butyl alcohol was added to 400 ml. of ammonia. Lithium (5.5 g., 0.79 mole) was added to the stirred solution over a two-hour period. The reaction mixture, which never became blue but developed a bronze-colored layer, was stirred for an additional four hours. Methanol ml.) was added and the ammonia was evaporated. Water and ether were added, the mixture shaken and the layers separated. The ether layer was washed twice with saturated sodium chloride solution. The aqueous layers were washed with a fresh portion of ether and the ether layers were combined, dried (Na S0 and concentrated by warming in vacuo. The syrupy residue was dissolved in 150 ml. of dioxane. The

dioxane solution was treated with ml. of 2N hydrochloric acid and heated in a nitrogen atmosphere on a steam bath for 30 minutes. Saturated sodium chloride solution and ether were added to the cooled reaction mixture, the mixture shaken and the layers separated. The ether was washed again with saturated sodium chloride solution. The aqueous layers were washed again with a fresh portion of ether. The combined ether layers were dried (Na SO and evaporated by warming in vacuo to leave 11 g. of an oily residue. Crystallization from acetone afforded 5.36 g. of 4,4aa,4 b,B,5,6,7,8,8aB,9,10-decahydro-7a-hydroxy-2(3H)- phenanthrone, m.p. 148-l51C., and a crop of 0.4 g., m.p. l44-l47C. Recrystallization from acetone provided a sample with m.p. l50152C.

EXAMPLE 9 4,4aa,4bB,5,6,7,8,8aa,9,10-Decahydro-7B-hydroxy- 2(3H)-phenanthrone [IX; 7B,4aa,4bB,8aoz-configuration] A solution of 18.33 g. (0.08 mole) 1,2,3 ,4,4aa,9, l 0, l 0aB-octahydro-7-methoxy-2aphenanthrol (VIIIB, Example 5) in 186 m1. of tetrahyadded, the mixture shaken and the layers separated.

The ether was washed with saturated sodium chloride solution and the aqueous layers were washed with a fresh portion of ether. The combined ether layers were dried (Na SO and the ether was removed by warming in vacuo. The remaining residue was taken up in 300 ml. of dioxane and 150 ml. of 2N hydrochloric acid. The solution was heated in a nitrogen atmosphere for 30 minutes on a steam bath. Saturated sodium chloride solution and ether were added to the cooled reaction mixture and the layers were separated. The ether was washed twice with saturated sodium chloride solution. The aqueous layers were washed with a fresh portion of ether. The combined ether was dried (Na SO and the solvent was removed while warming in vacuo, leaving 18 g. of an oily residue that crystallized from ether.

' afford Several recrystallizations from ether afforded 9.82 g. of 4,4aa 4l2 85,6.'7,8,8a0r,9, l -decahydro-7fi-hydroxy- 2(3H)-phenanthrone, rn.p. 141-143 C., when recrystailized from acetone.

EXAMPLE l0 4,4acz,4b,6,5,6,7,8,8aor,9,10-Decahydro-7a-hydroxy- 2(3ii)-phenanthrone UK; 7a.4-aa,4bfi,8aa-configuration1 l ,2,3,4,4aa1,2 ,10,1021,8-Octahydro-7-rnethoxy-2B- phenanthrol (VlllC, Example 6) (5.5 g., 0.024 mole), by the procedure of Example 9, afforded 2.8 g. of 4,4aa.4b,6.5.6,7,8,8a04,9,10-decahydro-7ot-hydroxy- 2(3l-i)-phenanthrone, m.p. 126.5128.5C., and a second crop of 0.6 g., mp. l23.5127C. (yield 64 percent), when recrystallized from acetone.

EXAMPLE 11 a.

l ,2,3,4,4acz,4l:-B,5,6,7,8,8a0z,9,10,10aa-Tetradecahydrophenanthrene-2,8,7/3-diol 7-acetate A solution of 6.95 g. (0.031 mole) 4,4a0t,4b.6,5,6,7,8,8a,8,9,10decahydro-7cv hydroxy- 2(3-l}-phenanthrone (Example 8) in 250 m1. of tetrahydrofuran and 250 ml. of ether was treated with ml. of dihydropyran and 200 mg. of p-toluenesulfonic acid, and the solution was left at room temperature overnight. Solid sodium carbonate was added to neutralize the acid and the solution was passed through a filter. Pyridine (2 ml.) was added and the volume of the solution was reduced to about ml. by heating the mixture in vacuo. Tetrahydrofuran (30 ml.) and absolute ethanol (60 ml.) were added to the residue and the resulting solution was added to 500 m1. of liquid ammo nia. A total. of 7 g.,(l mole) of lithium wire was added to the stirred ammoniacal solution over a 45 minute period. The solution remained blue for about one hour after addition of the lithium and 30 ml. more of absoiute ethanol was added to discharge the color. The volume reduced to one-third. Ether was added and the solution was heated under reflux conditions to remove more of the ammonia. Water was carefully added, followed by more ether. The layers were separated and the ether was washed with saturated sodium chloride sciatic-n. The aqueous layers were washed with a fresh portion of ether and the combined ether layers were dried (Na S0 The solvent was removed in vacuo to 1,2,3,4,4aez.4b[$,5,6,'7,8,8ao,9,10,10aoz-tetradecahydrophenanthrene-2BJ,G-diol 2dihydropyranyi ether. Pyridine (30 ml.) and acetic anhydride (15 mi.) were added to the residue. The next morning the reagents were removed by warming in vacuo. The residue was dissolved in 150 ml. of methanol and 150 ml. of 2N hydrochloric acid. At the end of 45 minutes, ether was added and the layers were separated. The ether was washed twice with saturated sodium chloride solution. The aqueous layers were washed i@aa-tetraclecahydrophenanthrene-2,B,7B-diol 7-acetate, mp. 166.5167C. when recrystallized from ether.

0. 1 ,2,3,4,4a0z,4bB,5,6,7,8,8aoz,9,10,10aoz-Tetradecahydrophenanthrene-2B,7B-diol A solution containing 150 mg. (0.6 mmoles) of 1,2,3 ,4,4aa,4bfi,5,6,7,8,8ao ,9, l 0,10aa-tetradecahydrophenanthrene-2,8,7B-diol.7-acetate in 15 m1. of 5- percent methanolic potassium hydroxide containing 5 1 percent water was boiled underreflux for 30 minutes. Water and ether were added, the mixture shaken and the layers separated. The aqueous layer was washed with a fresh portion of ether and the combined ether layers were washed with saturated sodium chloride, dried (Na SO and concentrated in vacuo. The residue crystallized from ether containing hexane and gave mg.(68 percent) of1,2,3,4,4aa,4bB,5,6,7,8,8aa,9,10,-

m.p. i

10aoz-tetradecahydrophenanthrene-2B,7B-diol, 152.5-153.5C. when recrystallized from acetone containing hexane.

(2.4 g., 0.01 mole) was added to 30 ml. of pyridine con-.

taining 2.4 g. of chromium trioxide. After standing for 24 hours at room temperature the reaction mixture was added to 500 ml. of ethyl acetate and then filtered. The solvent was removed in vacuo and ml. of ether was added to the residue. More salts were filtered away and the ether was evaporated, The residue containing 3 ,4,4act,4bB,5 ,6,7,8 ,8aa,9,10,10aa-dodecahydro-7B- hydroxy-2( ll-l)-phenanthrone 7-acetate was dissolved in 80 ml. of 5 percent methanolic potassium hydroxide containing 5 percent water and the solution was boiled under reflux for 45 minutes. Part of the methanol was removed by warming in vacuo. Ether and saturated sodium chloride were added. The mixture was shaken, the layers were separated and the ether waswashed with saturated sodium chloride solution. The aqueous layers were washed again with a fresh portion of ether and the combined ether layers were dried (Na SO and evaporated. Crystallization of theresidue from ether afforded 1.64 g. of 3,4,4a0z,4bB,5,6,7,8- 8am),10,1Oaat-dodecahydro-7B-hydroxy-2( ll-l)-phenanthrone, m.p. 1 l6117.5C. when recrystallized from ether.

EXAMPLE 13 3 ,4,4aa,4b,8,5 ,6,7,8,8a0z,9,10,10aB-Dodecahydro-7B- hydroxy-2( 1 H )-phenanthrone [X; 7B,4aa,4bB,8aa,1 OaB-configuration] dropping funnel used for the addition was rinsed with another 55 m1. of tetrahydrofuran. Ammonium chlo:

ride (10 g., 0.19 mole) was added as quickly as possible with controlled boilingLThe solvent was evaporated to half the volume. Water (300 ml.) and ether (300 ml.)

were added. The ether was boiled under reflux/to remove more of the ammonia. More ether and water were added and the layers were separated. The ether. layer was washed with saturated sodium chloride and was then dried (Na SO Evaporation of the ether left an oily residue of 11.6 g. The residue was chromatographed on 300 g. of silica gel. Elution with methylene dichloride-ether-pentane (225:3) afforded 7.9 g. of 3,4.4aa,4bB,5,6.7,8,8a0z,9,10.10aB-dodecahydro-7B- 17 hydroxy-2( lH)-phenanthrone, m.p. l07-108C. when recrystallized from acetone.

Further elution of the column with ether-methanol (19:1) afforded 0.9 g. of l,2,3,4,4aa,4b/3,5,6,7,8- ,8aa,9,10,10aB-tetradecahydrophenanthrene-Za, 7,8- diol, m.p. 216218C. when recrystallized from ether.

EXAMPLE 14 3,4,4aa,4bB,5,6,7,8,8aa,9,10,10aB-Dodecahydro-7ahydroxy-2(1l-l)-phenanthrone [X; 7a, 4aa,4bB,8aa, 1 OaB-configuration] Using the reduction procedure described in Example 13 4,4aa,4bB,5,6,7,8,8aa,9,1 -decahydro-7ahydroxy-2(3H)-phenanthrone (Example (3.2 g., 0.014 mole) afforded 1.49 g. of 3,4,4aa,4bfi,5,6,7,8- ,8aa,9, l O, 1 OaB-dodecahydro-7a-hydroxy-2( 1H )-phenanthrone, m.p. 137.5-138C. when recrystallized from ether.

3,4,4aa,4b,8,5,6,7,8,8aa,9,10,10aB-Dodecahydro- 7a-hydroxy-2(1H)-phenanthrone can be caused to react with methylmagnesium bromide to give 1 ,2,3,4,4ao ,4bB,5,6,7,8,8aa,9,10,lOaB-tetradecahydro-2-methyl-2,7a-dihydroxyphenanthrene, and the latter oxidized with chromic oxide to give 3,4,4aa,4bB,5,6,7,8,8aa,9,10,lOaB-dOdecahydro-7- methyl-7-hydroxy-2(1l-l)-phenanthrone [111; R and R" are H, X is H Z is (HO)(CH )C].

3,4,4aa,4bB,5,6,7,8,8aa,9,l0,lOaB-Dodecahydro- 7a-hydroxy-2( lH)-phenanthrone can be ketalized with ethylene glycol and the resulting ketal caused to react with sodium to give the sodio derivative of the 7ahydroxy group which in turn can be caused to react with methyl iodide to give the 7-methoxy compound. Hydrolysis of the ketal then gives 3,4,4aa,4bB,5,6,7,8- ,8aa,9,10,l0aB-dodecahydro-7a-methoxy-2(1H)-phenanthrone [111; R and R" are H, X is H Z is (a- CH O)CH].

' 3,4,4aa,4b/3,5,6,7,8,8aa,9,10,10aB-Dodecahydro- 7a-hydroxy-2(lH)-phenanthrone can be caused to .-react with p-toluenesulfonyl chloride in pyridine to give the 7a-p-toluenesulfonate, and the latter treated with anhydrous potassium fluoride in diethylene glycol, eighteen hours at 110C., to give 3,4,4aa,4bB,5,6,7,8- ,8aa,9,10,10aB-dodecahydro-7B-fluoro-2(1H)-phenanthrone [111; R and R" are H, X is H Z is (B- F)CH]. By analogous procedures the corresponding 7B-chloro and 7/3-bromo compounds can be prepared.

EXAMPLE l5 3,4,4aa,4bB,5,6,7,8,8aB,9,10,10aB-Dodecahydro-7ahydroxy-2( 1H )-phenanthrone [X; 7a,4aa,4bB,8aB, 1 OaB-configuration] Using the lithium-ammonia procedure of Example 13 for reduction, 4,4aa,4bB,5,6,7,8,8a/3,9,10-decahydro- 7a-hydroxy-2(3H)-phenanthrone (Example 8) (5.5 g., 0.025 mole) afforded 2.7 g. of 3,4,4aa,4bB,5,6,7,8- ,8aB,9,10,10aB-dodecahydro-7a-hydroxy-2( 1H )-phenanthrone, m.p. 8283C. when recrystallized from ether containing hexane.

EXAMPLE l6 3,4,4aa,4bl3,5,6,7,8,8aa,9, 10,1 OaB-Dodecahydro-7B- hydroxy-2( lH)-phenanthrone amidinohydrazone.

A solution of 3.5 g. (0.016 mole) of 3,4,4aa,4bB,5,6,7,8,8aa,9,l0,10aB-dodecahydro-7B- hydroxy-2(1l-l)-phenanthrone (Example 13) in 50 ml. of methanol was added to l 15 ml. of methanol containing 4.93 g. (0.036 mole) of aminoguanidine bicarbonate and 12 ml. of concentrated hydrochloric acid. After standing at room temperature overnight, the reaction mixture was added to 1 liter of ether. A precipitate formed which was found to be crude aminoguanidine hydrochloride and was removed by filtration. Dilution of the filtrate with more ether then precipitated the desired product (3.28 g., 66 percent). Recrystallization was effected by adding ether to a methanolic solution of the solid, to give 3,4,4aa,4bB,5,6,7,8,8aa,9,10,10aB

dodecahydro-7B-hydroxy-2( ll-l)-phenanthrone amidinohydrazone in the form of its hydrochloride salt, m.p. 245253C. (evacuated tube).

EXAMPLE 17 3,4,4aa,4bfl,5,6,7,8,8aa,9,10,10aB-Dodecahydro-7B- hydroxy- 2( 1H)-phenanthrone 2,2-ethylene dithioketal A solution of 4 g. (0.018 mole) of hydroxy ketone 3,4,4aa,4b,8,5,6,7,8,8aa,9,10,10aB-dodecahydro-7B- hydroxy-2(1H)-phenanthrone (Example 13) in 50 ml. of glacial acetic acid was treated with 5 ml. of ethane dithiol. The mixture was warmed slightly to effect solution and 4 ml. of boron trifluoride etherate was added to the warm solution. After the mixture had stood at room temperature for 5 minutes, water was added and the precipitated solid was collected. The solid (4.5 g.) was dissolved in ml. of 5 percent methanolic potassium hydroxide containing 5 percent water and the mixture was boiled under reflux for 30 minutes. The volume of methanol was reduced by warming in vacuo and ice water was added. The precipitate was recrystallized from acetone and afforded 2.94 g. of 3,4,4aa,4bB,5,6,7,8,8a0,9,10,1OaB-dodecahydro-7B- hydroxy-2( 1H)-phenanthrone 2,2-ethylene dithioketal, m.p. 129130C.

EXAMPLE 18 3,4,4aa,4bB,5,6,7,9,10,10a/3-Decahydro-7-oxo-2(1H)- phenanthrone [111; R and R" are H, X is l-1 ,Z is 0=C, A

A solution of 5.14 g. (0.023 mole) 4,4aa,4b/3,5,6,7,8,8aa,9,10-decahydro-7B-hydroxy- 2(3H)-phenanthrone (Example 9) in 50 ml. of pyridine was added to 50 ml. of pyridine containing 5.15 g. of chromium trioxide. After being stirred overnight the reaction mixture was added to 500 ml. of ethyl acetate. The mixture was filtered and the solvent was removed by warming in vacuo. Ether ml.) was added and more solid was filtered off. The solvent was evaporated leaving 4.69 g. of an oily residue that crystallized. Recrystallization from ether afforded 3.27 g. (63 percent) of 3,4,4aa,4bB,5,6,7,9,10,10aB-decahydro-7-oxo- 2(11-l)-phenanthrone, m.p. 124125C.

EXAMPLE 19 l,2,3,4,4aa,4bfl,5 ,6,7 ,8,8aa,9.10,10aB-Tetradecahydrophenanthrene-2,7-dione of l,2,3,4,4aa,4b,8,5,6,7,8,8aa,9,10,10aB-tetradecahydrophenanthrene-2,7-dione, m.p. 152.5l54C. Upon further concentration, another 0.7 g., m.p. 149.5-l51.5C. was obtained.

EXAMPLE 2O 1,2,3 ,4,4at,4b,8,5 ,6,'7,8,8a0t,9,l0,1OaB-Tetradecahydrophenanthrene-2,7-dione bisamidinohydrazone A solution of 4.83 g. (0.022 mole) 1,2,3,4,4aa,4bfi,5,6,7,8,8aa,9,10,10aB-tetradecahydrophenanthrene-2,7dione in 75 ml. of methanol was added to a solution of 13.6 g. (0.100 mole) of aminoquanidine bicarbonate and 35 ml. of concentrated hydrochloric acid in 315 ml. of methanol. After 24 hours at room temperature, the mixture was filtered to give 6.46 g. (73 percent) of l,2,3,4,4aoz,4bfi,5,6,7,8- ,8aa,9, l 0, l OaB-tetradecahydrophenanthrene-2,7- dione bisamidinohydrazone, mp. 350C.

EXAMPLE 21 3,4,4aoz,4b,6,5,6,7,8,8aa,9,lO,l0aB-Dodecahydro-7J- ethylenedimercapto-2( lH)-phenanthrone [111; R and R" are H, X is H Z is (ethylenedithio)C] A solution of 2.83 g. (9.4 mmoles) of 3 ,4,4aa,4b[3,5 ,6,7,8,8ao.',9,10,10aB-dodecahydro-7fihydroxy-2( ll-l)-phenanthrone 2,2-ethylene dithioketal (Example 17) in 40 ml. of pyridine was added to 40 ml. of pyridine containing 3 g. of chromium trioxide. After 5 days at room temperature the reaction mixture was added to 500 ml. of ethyl acetate. The mixture was filtered and the filtrate was concentrated to a residue by warming in vacuo. Ether (200 ml.) was added to the residue and more unwanted solid was separated. The ether was concentrated and 2 g. of 3,4,4aoz,4bfi,5,6,7,8,8a0z,9,lO,l0aB-dodecahydro-7,7- ethylenedirnercapto-2( 1H )-phenanthrone, mp. 13 l-132C. was obtained from the mother liquor (yield 76 percent).

EXAMPLE 22 EXAMPLE 23 lB,4bB-Dimethyl-3,4,4aa,4b,5,6,'7,8,8aa,9,10,10aB- dodecahydroJB-hydroxyQ(1H)-phenanthrone [111; R and R" are CH X is H Z is (B-HO)CH] Dry liquid ammonia was prepared by treatment of 300 ml. of liquid ammonia with lithium wire until the blue color persisted, followed by distillation until 100 ml. of liquid ammonia had been condensed in the reaction vessel. To the dry liquid ammonia was. added with stirring 0.16 g. (23 mmoles) oflithium wire followed by dropwise addition of a solution of 1.00 g. (4.29

mmoles) of 4,4a ,4b,5,6,7,8,8aot,9,lO-decahydro-7B- hydroxy-4bfl-methyl-2(3l-l)-phenanthrone in .15 ml. of tetrahydrofuran and 35 ml. of anhydrous ether over a 10 minute period. The mixture was stirred for 30 minutes and then treated with 1.2 ml. of methyl iodide which immediately discharged the blue color. The ammonia was allowed to evaporate and the residue was treated with ml. of water and extracted with 250 ml. of ether. The extract was washed with water and I saturated salt solution and dried (MgSOJThe residue (1.05 g), obtained upon evaporation of the solvent, was chromatographed on 30 g. of silica gel. Elution with 1:1 :3 methylene dichloride-etherpentane afforded crystalline material which, upon recrystallization from' acetonitrile, furnished 0.34 g. of product, m.p. 1 15120C. Recrystallization of the latter from acetonitrile gave 8aot9,10,1Oa,8-clodecahydr0-7;3-hydroxy-2(1lf l)-phenanthrone, mp. 129-132C.

EXAMPLE 24 The microorganism, Cunninghamella (American, Type Culture Collection No. 9244) was grown in surface growth 15 days old agar slants at 26C. containing as a nutrient medium:

(A) Maltese 40 gr. Proteose peptone 10 gr; Agar 15 gr. Distilled water to 1 liter,

and suspended in distilled water. lFive ml. portions of the suspension were used to inoculate 500 ml. Erlenmeyer flasks each containing 100 ml. of a sterilized medium (autoclaved at 15 lbs. per sq. in. for 20 minutes) I 7 containing:

(13) Cerelose (glucose) 5% Edamine (An enzymatic hydrolysate 2% of milk protein (Sheffield Farms Co.) Cornsteep 0.5% Tap water to 1 liter The seeded Erlenmeyer flasks were grown for, 72

hours at 26C. on a shaker rotating at 210 cycles per minute. 14-liter jar fermentors were prepared, each containing 10 liters of sterile medium B (sterilized for 45 minutes at 15 lbs. pressure) described above, and

seeded with 10 percent quantitites of inocula. The inoculated fermentors were agitated at 400 r.p.m. and

aerated with an air supply of 4 liters of air per. minute I Samples were taken from the tanks every 24 hours and analyzed chromatographically. Further incubation for 144 hours after addition of the'substrate indicated that this material had disappeared and the formation of.

three more polar metabolites was observed. The fermentation was terminated and the whole fermentation mash adjusted to pH 4.0 with 50 percent hydrochloric bainieri acid and extracted twice with 10-liter'portions of methylene dichloride. The extracts were combined and concentrated under reduced pressure to a residue. This residue was triturated twice with 250-ml. portions of n-pentane to remove some oils. The pentane washed fermentation extract (43 g.) from 21 g. of substrate was dissolved in ethyl acetate and put on 2 kg. of silica gel. The column was eluted with ethyl acetate. The first 8 liters eluted brown 'oils which were discarded. The next 7.5 liters of ethyl acetate eluted 4.38 g. of a mixture of hydroxylated products. Continued elution with ethyl acetate afforded 5.98 g. of oil containing mostly the more polar 4,4aa,4bB,5,6,7,8,8aa-9,l-decahydro- 78,10a-dihydroxy-2(3l-1)-phenanthrone. After preparative thick layer chromatography of these two fractions 1.45 g. of 4,4aa,4bB ,5,6,7,8,8aa,9,lO-decahydro- 713,10a-dihydroxy-2(3H)-phenanthrone was obtained, m.p. 143l46C. Recrystallization from acetone containing ether gave a sample with m.p. 155l56C.

From the less polar band 1 g.

l ,2,3,4,4aa,4b 8,5,6,7,9,10,10aB-dodecahydro-7- oxophenanthrone-2a,4bB-diol, m.p. l5ll54C. was obtained. This material was recrystallized from acetone containing ether to give a sample with m.p. 163l65C. indistinguishable by infrared analysis, TLC analysis and mixed melting point from a sample prepared by an alternative chemical method as described below.

By substituting as the substrate in the foregoing preparation, 4,4aa,4b,5,6,7,8,8aa,9,10-decahydro-7B- hydroxy-4bB-methyl-2(1H)-phenanthrone (X1), there can be obtained 4,4aa,4b,5,6,7,8,8aa,9,lO-decahydro- 4bB-methyl-7B, 1 0a-dihydroxy-2(3 H )-phenanthrone.

In the foregoing fermentation process, the organism Cunninghamella bainieri can be replaced by Cunninghamella verticillator or Cunninghamella elegans.

EXAMPLE 25 a. l ,2,3,4,4aa,5,6,7,8,9,10,10aB-Dodecahydro-7-oxo-2aphenanthrol A solution of 13.8 g. (0.06 mole) of 1 ,2,3,4,4aa,9,10,10aa-octahydro-7-methoxy-2B- phenanthrol and Jar-phenanthrol in 50 ml. of tetrahydrofuran was added to 300 ml. ofliquid ammonia. Lithium wire (4.7 g., 0.66 mole) was added over a half-hour period with stirring. After stirring the mixture for a halfhour longer, a mixture of 70 ml. of absolute ethanol and 70 ml. of ether (anhydrous) was added to discharge the blue color. The solution was evaporated to half the volume. Ether and water were added and, after shaking, the layers were separated. The ether was washed with saturated sodium chloride solution and the aqueous layers were washed with a fresh portion of ether. The combined ether layers were dried (Na SO and the ether was removed by warming in vacuo. The remaining residue was taken up in 100 ml. of tetrahydrofuran and 200 ml. of methanol. Oxalic acid (10 g.) in 75 ml. of water was added. The solution was kept at room temperature for 75 minutes and then added to a large volume of ether. The layers were separated and the ether layer was washed several times with saturated sodium bicarbonate solution. The aqueous layers were washed with a fresh portion of ether. The combined ether layers were dried (Na SO and the solvent was removed by warming in vacuo. The residue crystallized from ether containing hexane and afforded 7.9 g. of

1,2,3 ,4,4aa,5,6,7,8,9, l O, l OaB-dodecahydro-7-oxo-2aphenanthrol, m.p. 123C.

A solution of 6.76 g. of 85 percent m-chloroperbenzoic acid in 80 ml. of methylene dichloride was added to 1,2,3,4,4aa,5,6,7,8,9,10,l0aB-dodecahydro-7-oxo- Zen-phenanthrol (6.5 g., 0.29 mole) in 50 ml. of methylene dichloride. After standing at room temperature overnight, the reaction mixture was washed twice with saturated sodium bicarbonate solution and once with saturated sodium chloride solution. The aqueous layers were washed again with methylene dichloride. The organic layers were combined and dried with sodium sulfate. The solvent was removed by warming in vacuo, leaving 6.3 g. of an oily residue that would not crystallize. The residue was chromatographed on 500 g. of silica gel. Ethyl acetate with 0.5 percent methanol was used for elution. A less polar fraction (2.53 g.) was obtained which on crystallization and recrystallization from acetone containing hexane afforded 1.43 g. of

1,2,3,4,4aa,4b,5,6,7,9,10,10aB-dodecahydro-7-oxophenanthrene-2oz,4bB-diol, m.p. -l 61C.

A more polar fraction (1.74 g.) afforded, on crystallization and recrystallization from acetone-hexane, 0.88 g. of 1,2,3,4,4acr,4b,5,6,7,9,10,10aB-dodecahydro-7- oxo-phenanthrene-2a,4ba-diol, m.p. 176-177C.

EXAMPLE 26 l,2,3,4,4aa,4b,8,5,6,7,8,8aa,9,10, 1 OaB-Tetradecahydro-7B-acetoxy-phenanthrene-2,lO-dione [111; R and R" are H, X is 0, Z is (B-CH COO)CH] A solution of 900 mg. of 4,4aa,4b,8,5,6,7,8,8aa,9,10- decahydro-7B,10ct-dihydroxy-2(3H)-phenanthrone (Example 24) in 50 ml. of acetic acid and 5 m1. of 10 percent sulfuric acid was kept at room temperature in a nitrogen atmosphere for 24 hours. At the end of this time the solution was poured into ice and solid sodium bicarbonate was added. Ether was added and the layers were separated. The ether was washed with saturated sodium bicarbonate solution and then with saturated sodium chloride solution. The aqueous layers were washed with more ether. The ether was combined and dried over sodium sulfate. The ether was evaporated by warming in vaco leaving a residue that partially crystallized. Ether containing hexane was added and 310 mg. of l,2,3,4,4a0z,4bfi,5,6,7,8,8aa,9,10,10aB-tetradecahydr0-7B-acetoxyphenanthrene-Z,lO-dione. Recrystallization from ethergave a sample with m.p. 146-148C.

EXAMPLE 27 ,3,4,4aa,4b,5,6,7,9,10,10afi-Dodecahydro-4b6 -hydroxyphenanthrene--2,7-dione [111; R is OH, R is 1-1, X is H Z is O=C, A

A solution of 1,2,3,4,4aa,4b,5,6.7,9,l0,IOaB- dodecahydro-7-oxophenanthrene-2a,4bB-diol (Example 24) in 35 ml. of pyridine was added to 35 ml. of pyridine containing 3 g. of chromium trioxide. After standing at room temperature overnight, the reaction mixture was diluted with a large volume of ethyl acetate and filtered. The solvent was removed by warming in vacuo. The residue was taken up in acetone ether and was filtered. The volume was reduced and there was obtained 2 g. of 1,2,3,4,4aa,4b,5,6,7,9,10,l0a,B-

23 clodecahydro-4b/3-hydroxy-phenanthretie-2,7-dione. mp. 179180C. when recrystallized from ether.

EXAMPLE 28 a. 3,4,4acx,4b,5,6,7,8,8aa,9,l0,10aB-DodecahydroJB- hydroxy-4bB-methyl-2(1H)-phenanthrone 7-p-toluenesulfonate concentrated to a residue to give 16.4 g. of 3,4,4a,4b,5,6,7,8,8aa.9,10,10afi-dodecahydro-7flhydroxy-4ofl-methyl-2( l H )-phenanthrone 7-ptoluenesult'onate as an amber oil. This oil was used without further purification in the following procedure.

b. Detosylation of 3.4,4aor,4b.5.6,7,8,8aa,9,l0,10aB-dodecahydro-7B- hydroxy-4hfi-methyl-2( 1 H )-phenanthrone 7-p-toluenesulfonate A solution of 16.3 g. of the above oily tosylate in 100 m1. of s-collidine was heated under reflux for 4 hours. The solution was cooled and poured into 400 ml. of 2N sulfuric acid. The acid mixture was extracted with ether and the extract was washed with 2N sulfuric acid, water and oririe. It was dried (MgSO and concentrated to a residue to give 7.0 g. of an oil which was assumed to he a mixture of A and N-Z-phenanthrones. This mixture was hydrogenated in the fo lowing procedure without purification.

c. Hydrogenation of the mixture of A and N-Z-phenanthrones.

A solution of 0.69 g. of the mixture of phenanthrones described in the preceding example in 25 ml. of undenatured ethanol was treated with 0.10 g. of percent palladium-on-charcoal and the mixture was shaken in a hydrogen atmosphere until one molar equivalent was absorbed (5 minutes). The mixture was filtered and the filtrate was concentrated to a residue by warming under reduced pressure. The residue was dissolved in ether and the solution was dried (MgSO and concentrated to give 0.56 g. of 3,4,4aa,4b,5,6,7,8,8aa,9,l0,- ll0a,8d0decahydro-4bfi-methyl-2( 1H )-phenanthrone fill; R is CH R" is H, X is H Z is CH as an amber oil. it was purified by column chromatography on g.

' of silica gel with elution by 1:9 ether-pentane.

EXAMPLE 29 a. 8,8-Dimethyl-l,2,3,4,4a0z,4b}3,5,6,7,8,l0,IOaB- dodecahydro-7-oxo-2a-phenanthrol [XV; R is H, lower-alkyl is CH over a half-hour period. The solution wascstirred at room temperature for 1 hour and was finally heated V under reflux for 10 minutes. The reaction mixture was cooled anddilute hydrochloric acid was added. Ether and water were added and the layers were separated. The ether was washed with dilute sodium hydroxide.

and then saturated sodium chloride solution. Theether was dried (Na SO and evaporated to afford an oily residue that crystallized from ether containing hexane. Recrystallization twice from the same solvent gave 1 1.0 g. of 8,8-dimethyl-1,2,3,4,4aa,4b5,5,6,7,8,l0,10aB- dodecahydro-7-oxo-2a-phenanthrol, m.p. 1163-4 18C.

Upon concentration of the mother liquor another crop of 3.2 g., mp. 114-116C. was obtained. The mother (225:3) afforded another 5 percent). A sample recrystallized from ether ing hexane melted 1Zi122.

b. 3 ,4,4aaa,4bfi,5 ,6,7,8, 10,10aB-Decahydro-S.S-dirnethyl- 7-0xo-2(lH)-phenanthrone [XVI; R is H, lower-alley] is CH A solution. of 16.6 g. (0.067 mole) of the hydroxy keto'ee of part (a) in 210 ml. of pyrit,.,\rie was addedto 210 ml. of pyridine containing 16.6 g. (0.166 mole) ofchromium trioxide. The mixture was stirred overnight and was then added to about ml. of ethyl acetate. The

mixture was filt red through infusorial earth and the soivent was re: ed by warming in vacuo. Ether was added to the residue and more inorganic solids were filtered away. The ether was concentrated to afford 11.9. g. of 3,4,4aa,4h 6,5,.6,7,8,l0,i0aB-decahydro-8,8-

dimethyl-7-ox -2( Hi-phena r ap.

13="--133C. A so 1.21. ,g., 128---13C-C. was c ed. mother liqnorwas chromatographed on 15: g. of silica gel. Elution with methylene dichlcride-ether-pentane (2:315) afforded, 7

another 0.7 g., mp. l31l3?iC. (33 p cent). A sari ple recrystallized i'rorn ether melted at 13F -134C Similarly, starting from l,2,3,4.,4aa,4h,5 decahvdro-2m-hydroxy-4h,8-methyl-7(5H)-pherianthrone [Xi\/; R. is CH there he prepared 3,4.4aa,4b,5,6,7,8,10,lQafi-decahydro-lhflfifitrimethyl-7-oxo-2( ll-l)-phenanthrorie IXVI; R is CH lower-alkyl is CH 3.4,4a.rx,4h,5,6,7,8,10,l0a,8- Decahydro-4bB,8 .8-trimethyi-?-oxo-2( 1H )-phenanthrone reacts with e hylene glycol in the preserice of p-toluenesulfonic acid give the 2-mono-eti1ylene glycol ketal. The latter ketal is reduced with lithium aluminum hydride to give decahydro-4hfi,8,8-trimethyl-7;3-hydroxy-2( 1H phenanthrone 2-monoethylene glycol ketal. which is further-reduced catalytically in the presence of, palladiurrif on-carbon to give 3,4,4aa,4h,5,6,7,3,8aoi,9;i0,10afi- 2-mono-ethylerie I I EXAMPLE 30 a. Mixture of 10aand lOB-acetoxy-8,8dimethyl-3,4,4aa,4bfi,5 ,6,7,8, 1 O,- laB-decahydro-7-oxo-2( l H)-phenanthrones cuprous chloride were added. After the reaction mixture was heated under reflux for another 7 hours, the above addition procedure was repeated. The reaction mixture was then heated under reflux for 24 hours. The reaction mixture was added to ice water. Ether was added and the layers were separated. The ether was washed with saturated sodium bicarbonate and satu rated sodium chloride solutions and then dried (Na S0 Evaporation of the solvent afforded an oily residue that was redissolved in ether and filtered. Evaporation of the solvent by warming in vacuo gave 190 mg. of an oily residue. This residue was chromatographed on 19 g. of silica gel. Elution with 400 ml. of methylene dichloride-ether-pentane (2:315) gave 50 mg. (20 percent) of starting diketone. Continued elution with another 60 ml. of the same solvent mixture afforded 60 mg. (20 percent) of the mixture of 1004- and IOB-acetates.

b. Mixture of 10aand l0B-acetoxy-8,8-dimethyl-l ,2,3,4,4aa,4bB,- 5 ,6,7 ,8, l 0, l OaB-dodecahydro-Za-phenanthrol.

A solution of 2.3 g. (9.3 millimoles) of 8,8-dimethyl- 1,2,3 ,4,4aa,4bB,5,6,7,8, l 0, l OaB-dodecahydro-7-oxo- Za-phenanthro] in 50 ml. of benzene and 5 ml. of acetic acid was treated with 1.7 ml. (10.0 millimoles) of tbutyl peracetate (75 percent in benzene). Cuprous chloride (10 mg.) was added and the reaction flask after being flushed with nitrogen was boiled under reflux in a nitrogen atmosphere for 24 hours. Another 2 ml. of t-butyl peracetate was added and the reaction mixture was boiled under reflux for another 24 hours. The reaction mixture was worked up as described above in part (a). The oily residue (2.65 g.) was chromatographed on 200 g. of silica gel. Elution with methylene dichloride-ether-pentane (2:3:5) afforded 540 mg. (23 percent) of starting material. Continued elution with methylene dichloride-ether (1:4) then gave 690 mg. (19 percent) of the mixture of acetates.

EXAMPLE 31 8,8-Dimethy1-3,4,4aa,4bB,5,6,7,8,l0,10aB-Decahydro-7,l0-dioxo-2(lH)-phenanthrone [XVI]; R is H, lower-alkyl is CH;,]

a. The mixture of acetates from parts (a) and (b) of Example 30 were combined and were dissolved in 30 ml. of 5 percent methanolic potassium hydroxide containing 5 percent water. The solution was boiled under reflux in a nitrogen atmosphere for 45 minutes. The reaction mixture was added to ice and water. Sodium chloride was added followed by ether. The layers were separated. The ether was washed twice with saturated sodium chloride solution. The aqueous layers were washed with a fresh portion of ether and the combined ether layers were then dried (Na SO and evaporated by warming in vacuo to afford 390 mg. of an oily residue. The residue, containing a mixture of 1004- and lOB-hydroxy-S,8-dimethyl-3,4,4aa,4bB,5.6,7,8,l0,- l0aB-decahydro-7-oxo-2( 1H )-phenanthrones and 10aand lOB-hydroxy-8,8-dimethyll,2,3,4,4aa.4bfi.5,6,7,8,l0,10aB-dodecahydro-2aphenanthrols was dissolved in 20 ml. of pyridine and this solution was added to 20 ml. of pyridine containing 600 mg. of chromium trioxide. The reaction mixture was kept at room temperature for 40 hours and then added to about ml. of ethyl acetate. The mixture was filtered and warmed in vacuo to remove the solvent. Ether was added and more inorganic solids were filtered away. The ether was evaporated. The residue was crystallized from ether and afforded 140 mg. of 8,8-dimethyl-3,4,4aoz,4bB,5,6,7,8,10,10aB-decahydro- 7,10-dioxo-2(lH)-phenanthrone, m.p. l47-l50C. (23 percent). Recrystallization from ether gave a sample melting at l52-l53C.

b. A solution of 19 ml. of t-butyl chromate in carbon tetrachloride, 20 ml. of carbon tetrachloride, 6 ml. of acetic acid and 2.5 ml. of acetic anhydride were heated at 50C. in a water bath. A slow stream of air was blown through the solution while it was being stirred. A solution of 1 g. (4.1 millimoles) of3,4,4aa,4bfl,5,6,7,8,10,- 10aB-decahydro-8,8-dimethyl-7-oxo-2( lH)-phenanthrone in 18 ml. of carbon tetrachloride was added during a half-hour period. The temperature was maintained at 50-70C. for a 2 hour period. This solution was stirred at room temperature with a slot stream of air passing through for another 16 hours. Carbon tetrachloride (50 ml.) was added to the reaction mixture (to maintain volume). Oxalic acid (7.5 g., 83 millimoles) in 75 ml. of water was added during a half-hour period while the solution was being stirred in an ice bath. Oxalic acid (5.25 g.) was again added and the reaction mixture was stirred for another two hours. More carbon tetrachloride and water were added and the layers were separated. The aqueous layer was washed with a fresh portion of carbon tetrachloride. The carbon tetrachloride solutions were combined and were washed twice with saturated sodium bicarbonate and once with saturated sodium chloride solution. The organic solution was dried (Na SO and the solvent was removed by warming in vacuo to afford 320 mg. of a residue. The aqueous layers were extracted with ethyl acetate and gave another 570 mg. of oily residue upon evaporation. Crystallization from ether afforded 225 mg. of 8,8-dimethyl-3,4,4aa,4bB,5,6,7,8,10,10aB-decahydro- 7,l0-dioxo-2( lH)-phenanthrone, m.p. 148151C.

EXAMPLE 32 Ethyl 3,4,4aa,4bB,5,6,7,8,8aa,9,10,10aB-dodecahydro-7B- hydroxy-A -phenanthreneacetate (mixture of isomers) [1; R is C H R, R and R are H, X is H Z is (B-HO)CH] A solution of sodium ethoxide [prepared from 1.6 g. (0.070 mole) of sodium and absolute ethanol] in ml. of dry dimethylformamide (DMF) was cooled in an ice bath and treated dropwise with a solution of 15.5 g. (0.070 mole) of triethyl phosphonoacetate in 20 ml. of dry DMF with stirring. The resulting solution was stirred cold (0C.) for 5 minutes and then a solution of 7.68 g. (0.0346 mole) of 7,8-hydroxy' 3,4,4aa.4b 3,5,6,7,8,8aa,9,l0,10aB-dodecahydro- 2(ll-i-)-phenanthtone (Example 13) in 30 ml. of dry DMF was added dropwise with stirring. This mixture was then stirred cold (C.) for minutes and at room temperature for 2 hours. it was added to 1.5 liter of water and the mixture was made acidic with 2N hydrochloric acid. The precipitated product was extracted with ether and the extracts were washed with brine and dried over sodium sulfate. Removal of the ether gave an oily residue which partially crystallized upon addition of about 25 ml. of ethanol. Dilution of this mixture with 300 ml. of water and filtration afforded 10.4 g. of a crystalline product mp. 8910lC., which was shown by gas-liquid phase chromatography (glpc) to be a 1:1 mixture of cis and trans isomers together with 1.6 percent of an impurity. This mixture was purified on silica zhromatoplates developed with pure ether and then recrystallized from ether-hexane to give ethyl 3,4,4act,4b,8,5,6,7,8,8aot,9,10,10aB-dodecahydro-7B- hydroxyA 0! -phenanthreneacetate, m.p. l02-1 10C.

EXAMPLE 33 Methyl l ,8,4bB-dimethyl-3,4,4aa,4b,5,6,7,8,8aoz,9,10,10a,8- clodecahydro-7/3-hydroxyA -phenanthreneacetate ll; R is CH R is H, R and R" are CH X is H Z is (B-HO)CH] To a suspension of 2.82 g. (0.052 mole) of reagent sodium methoxide in 35 ml. of dry 1,2-dimethoxyethane was added 9.5 g. (0.052 mole) of trimethyl phosphonoacetate in 35 ml. of dry 1,2-dimethoxyethane and the mixture was stirred for 1 hour at room temperature. A solution of 6.5 g. (0.026 mole) of 15,4bfidimethyl-7fi-hydroxy-3 ,4-,4a0z,4b,5 ,6,7,8 ,8a0z,9, l 0,- 10afi-dodecahydro-2(ll-l)-phenanthrone (Example 23) in 70 ml. of 1,2-dimethoxyethane was added. Silica plates developed with methanol-ether (1:49) showed that the reaction was about 50 percent complete in one hour but that it progressed no further even when the mixture was refluxed (84C.) for five days. After the reflux period, 25 ml. of water and 500 ml. of ether were added and the layers were separated. The organic layer was washed with brine, dried over magnesium sulfate and concentrated to give 9.0 g. of methyl 1,8,4bl3- dimethyl-3,4,4aa,4b,5 ,6,7,8,8aot,-9,l0,10a,8- dodecahydro-7B-hydroxy-A -phenanthreneacetate as a viscous oil which contained considerable unreacted ketone. This mixture was hydrolyzed directly, giving a carboxylic acid which was easily separable from ketonic impurity.

EXAMPLE 34 A solution of 25.4 g. (0.087 mole) of ethyl 3,4,4aa- 4bl3,5,6,7,8,8aa,9,10,10aB-dodecahydro-7B-hydroxy- A -phenanthreneacetate (Example 32) in 220 m1. of pyridine was added in 2 minutes with stirring to a mixture of 21.9 g. (0.22 mole) of chromium trioxide and 220 ml. of pyridine at room temperature and the resulting mixture was stirred overnight. Ethyl acetate (1.5 liter) was added, the mixture was filtered and the filtrate was concentrated to a residue by warming under reduced pressure. This residue was treated with 400 ml. of ether and further insoluble material was rei moved by filtration. Concentration of the ether solution and addition of hexane afforded 6.13g. of ethyl 3,4,4arx,4bB,5,6,7,8,8aoz.9,10,10a/3-dodecahydro-7- oxo-Zt -phenanthreneacetate (isomer A), mp. 96.5C. when recrystallized from ether-hexane.

b. 3,4,4a0t,4b 8,5,6,7,8,8aor,9,l0,10afi-Dodecahydro-7- oxo-d -phenanthreneacetic acid (llsomer B) [l; R, R", R and R are H, X is H Z is O=C1 The mother liquor residues from the preceding experiment containing ethyl 3,4,4aot,4bfi,5,6,7,8,8-

reneacetate were dissolved in 500 ml. of 95 percent ethanol, 200 ml. of 2N aqueous sodium hydroxide was added and the solution was refluxed in a nitrogen atmo sphere for 75 minuteshThe reaction mixture was added to ice-water, neutralized with acetic acid and the prod-,

8.22 g. (0.0313 mole) ofa mixture of isomer A and Isomer Bunsaturated carboxylic acids. This solid, mp. 180-195C., was dissolved in 250 ml. methanol, 0.l0

mole of diazomethane in ether was added and the solution was allowed to stand overnight. The solvent was removed and the crystalline residue was recrystallized from ether by the addition of hexane to give 2.87 g. of

material which melted at l00130C. Two further recrystallizations furnished 1.6 g. I of 3,4,4a0z,4bfi,5,6,7,8,8a0:,9,l0,l0a;8-dodecahydro-7- OXO-AZQH) -phenanthreneacetate (isomer B), m.p.'

EXAMPLE 35 dro-7fi-hydroxy-Z-phenanthreneacetate (isomer A) [1;

R is C ll lR, R and R" are H, X is H Z is (fi-HOKIH, saturated side chain] A solution of 12.0 g. (0.041 mole) of ethyl 3,4,4aot,4bfl.5,6,7,8,8aa,9,l0,10aB-dodecahydro-7B- hydroxy-A -phenanthreneacetate (Example 32) in 300 ml. of absolute ethanol was hydrogenated at 3.9 l g./cm and 25C. for two and one-half hours in the presence of 1.2 g. of 10 percent palladium-on-carbon. The mixture was filtered and the filtrate was concentrated to a residue by warming under reduced pressure.

The residue was dissolved in ether and precipitated by addition of a small amountof hexane to give 4.43 g. of material, m.p. 1 l0-l C. Concentration of the filtrate gave 1.07 g. of material, mp. 106l08C. (41 percent). Recrystallization from ether containing hexane afforded ethyl l,2,3,4,4aa,4b,8,5,6,7,8,8aa,9,10,

Methyl methyl i 29 lOaB-tetradecahydro-7B-hydroxy-2-phenanthreneacetate (lsomer A), mp. l-l l 1C.

The mother liquor residues were greatly enriched in Isomer B but TLC showed no separation of these isomers when using silica plates developed with 100 percent ether or 3:7 pentane-ether. These residues were hydrolyzed as described below in Example 53.

By the foregoing procedures the following examples of lower-alkyl esters (I; R is lower-alkyl) were prepared. In most instances the esters were not purified but were hydrolyzed directly to the corresponding free acids as described below.

EXAMPLE 36 Methyl 3,4,4aa,4b,5,6,7,8,8aa,9,10,10aB-dodecahydro-4bB-methyl-A -phenanthreneacetate [1; R is CH R is H, R is CH R" is H, X is H Z is CH ]from 3,4,4aa,4b,5,6,7,8,8aa,9,10,10aB-dodecahydro-4bB- methyl-2( l H)-phenanthrone (Example 28) and trimethyl phosphonoacetate.

EXAMPLE 3 7 Methyl 3,4,4aa,4bB,5,6,7,8,8aa,9,l 0,10aB- dodecahydro-7B-hydroxy-A -phenanthreneacetate [1; R is CH R, R and R" are H, X is H Z is (B- HO)CH], from 7B-hydroxy-3,4,4aa,4bB,5,6,7,8- ,8aa,9,l0,l0a,B-dodecahydro-2( lH)-phenanthrone (Example 13) and trimethyl phosphonoacetate.

EXAMPLE 38 Ethyl 3,4,4aa,4bB,5,6,7,8,8aa,9,10,10aB-dodecahydro-7oz-hydroxy-A -phenanthreneacetate [1; R is C 11 R, R' and R are H, Z is (a-HO)CH], from 3,4,4aa,4b/3,5,6,7,8,8aa,9,10,10aB-dodecahydro-7ahydroxy-2(lH)-phenanthrone (Example 14) and triethyl phosphonoacetate.

EXAMPLE 39 Methyl 3,4,4aa,4bB,5,6,7,9,10,10aB-decahydro-7- oxo-A a -phenanthreneacetate [1; R is C 11 R,R and R are H, X is H Z is O=C, A from 3,4,4aa,4bB,5 ,6,7,9,l0,10aB-decahydro-7-oxo-2(1H)- phenanthrone (Example 18) and trimethyl phosphonoacetate.

EXAMPLE 40 Methyl 3,4,4aa,4bB,5,6,7,8,8aa,9,10,10aB- dodecahydro-7,7rethylenedimercapto-A phenanthreneacetate [1; R is CH R, R and R" are H X is H Z is ethylenedithio], from 3,4,4aa,4bl3,5,6,7,8- 5O ,8aa,9,l0,10aB-dodecahydro-7,7-ethylenedimercapto- 2(lH)-phenanthrone (Example 21) and trimethyl phosphonoacetate.

EXAMPLE 41 Methyl 3,4,4aa,4b,5,6,7,8,8aa,9,l0,IOaB-dodecahydro-4bB-methyl-7B-hydroxy-A -phenanthreneacetate [1; R is CH R is H, R is CH R" is H, X is H Z is (B-HO)CH], from 3,4,4aa,4b,5,6,7,8- ,8aa,9,l0,10aB-dodecahydro-7B-hydroxy-4bB-methyl- 2( lH)-phenanthrone (Example 22) and trimethyl phosphonoacetate.

EXAMPLE 42 Methyl 3,4,4aa,4b,5,6,7,8,8aa,9,l0,lOaB-dodecahydro-4bB-methyl-7-oxo-A -phenanthreneacetate [1; R is CH R is H, R is CH R is H, X is H Z is O=C], by oxidation of methyl 3,4,4aa,4b,5,6,7,8-

39 ,8aa,9,lO,lOaB-dodecahydro-4bB-methyl-7B-hydroxy- A -phenanthreneacetate (Example 4]).

EXAMPLE 43 Ethyl 3,4,4aa,4b,5,6,7,8,8aa,9,l0,10aB-dodecahy-' eacetate [1; R is C H R is H, R is CH R is H, X is H Z is (B-HO)CH], from 3,4,4aa,4b,5,6,7,8- ,8aa,9,l0,lOaB-dodecahydro-7B-hydroxy-4bB-methyl- 2( lH)-phenanthrone (Example 22) and triethyl phosphonoacetate.

EXAMPLE 44 Ethyl 3,4,4aa,4b,5,6,7,8,8aa,9,l0,10aB-dodecahydro-4bB-methyl-7-0xo-A -phenanthreneacetate [1; R is C 11 R is H, R is CH R is 1-1,-X is H Z is O=C], by oxidation of ethyl 3,4,4aa,4b,5,6,7,8- ,8aa,9,10,10aB-dodecahydro-4bB-methyl-7B-hydroxy- A a -phenanthreneacetate (Example 43).

EXAMPLE 45 Ethyl 3,4,4aa,4b,8,5,6,7,8,8aB,9,l0,10aB-dodecahydro-7a-hydroxy-A -phenanthreneacetate [1; R is C H R, R and R are H, X is H Z is (a-HO)CH, A/B cis], from 3,4,4aa,4bfi,5,6,7,8,8aB,9,10,10a,8- dodecahydro-7a-hydroxy-2(lH)-phenanthrone (Example 15) and triethyl phosphonacetate.

EXAMPLE 46 Ethyl 3,4,4aa,4b,8,5,6,7,8,8aB,9,lO,lOaB-dodecahydro-7-oco-A -phenanthreneacetate [1; R is C2H oxidation of ethyl 3,4,4aa,4b,8,5,6,7,8,8aB,9,10,10a/3- dodecahydro-7oz-hydroXy-A -phenanthreneacetate (Example 45).

EXAMPLE 47 Methyl 3,4,4aa,4bB,5,6,7,8,8aa,9,10,10aadodecahydro-7B-hydroxy-A -phenanthreneacetate [1; R is CH R, R and R are H, X is H Z is (B- HO)CH, B/C cis], from 3,4,4aa,4bB,5,6,7,8,8aoz,9,l0,- 10aa-dodecahydro-7,B-hydroxy-2( 1H )-phenanthrone (Example 12) and trimethyl phosphonoacetate.

EXAMPLE 48 Methyl 3,4,4aa,4b,5,6,7,9,l0,l0aB-decahydro-4B- liydroxy-7-oxo-A -phenanthr'eneacetate [1; R is CH R is H, R is OH, R is H, X is H Z is O=C, A m.p. l621 96C. (from ether), from 1,2,3,4,4aa,4b,5,6,7,9,10,10aB-dodecahydro-4bB- hydroxyphenanthrene-Z,7-dione (Example 27) and trimethyl phosphonoacetate.

EXAMPLE 49 EXAMPLE Ethyl 3 ,4,4,aa,9, l O, l OaB-hexahydro-7-methoxy- A -phenanthreneacetate [11; R is C H R is H, R" is CH from 3,4,4aa,9,lO,lOa,8-hexahydro-7- Sill methoxy-2(lH)-phenanthrone (Example 7) and triethyl phosphonoacetate.

EXAMPLE 51 Methyl 8,8-dimethyl-7,lO-dioxo- 3,4,4a:,4b,8,5,6,7,8,iO,i()a-decahydro-A phenanthreneacetate [l; R is CH R, R and R are H, X is 0, Z is O' C, 8,8-(Ci'i LX isomer A, m.p. i42-143 C. (from ether); and isomer B, m.p. 123-l24C (from ether), from 3,4,4-atx,4bfl,5,6,7,8,10,10aB-decahydro-S,E-dimethyl- 7,lO-dioxo-2(lH)-phenanthrone and trimethyl phosphonoacetate.

EXAMPLE 51A Ethyl 3,4,4aa,4b,5,6,7,8,8a0z,9,10,iOa,8-dodecahydro-4b/B-n1ethyl-7Bhydroxy-A -phenanthrenepropionate [1; R is C H R and R are CH R is H, X is H Z is (B-HO)CH], from 3,4,4aa,4b,5,6,7,8- ,iiacz,9,1 0, lOaB-dodecahydro-4bB-methyl-7B-hydroxy- 2( liiyphenanthrone and triethyl oc-phosphonopropionate.

EAAMRLE 51B Methyl 3,4,4a0,4b,5,6,'7,8,8ac4,9,lO,l0afi-dodecahydroibfii;8,8-trimethyl7fi-hydroxy-N -phenanthreneacetate [i; R is CH R" is H, R is CH R" is H, X is H Z is (fl-HOMIH, 8,8-(CH from 3 ,4,4-ao4,4-o,5 ,6,7,3,8acz,9,10,10aB-dodecahydro- 4bfi,8,8-trimethyl-7B-hydroxy-2( 1H)-phenanthrone and trimethyl phosphonoacetate.

According to the procedures of Examples 32 and 33, the following compounds can be reacted with trimethyl phosphonoacetate:

3 ,4,4a0z,9,1O,1Oaoz-Hexahydro-"l-rnethoxy-2( 1H)- phenanthrone (Example 4),

3,4,4aa,4b,8,5,6,7,8,8a01,9,lO,lOaBDodecahydro-7- iethyl-7-hydroxy-2( 1H )-phenanthrone,

3,4,4a0r,4bfi,5,6,7,8,8a0z,9,lO,l0a,8-Dodecahydro- 7oi-methoxy-2( 1H )-phenanthrone,

3,4,4acz,4b,8,5,6,7,8,8av,9,10,10afi-Dodecahydro- 7,6-fluoro-2(lH)-phenanthrone,

3,4,4aoz,4b,6,5,6,7,8,8aoi,9,lO,10a,8-Dodecahydro- "/fi-chloro-2( 1H )-phenanthrone,

3,4,4a0z,4bfi,5,6,7,8,8aoz,9,10,1OaB-Dodecahydro- '7{3br0m0-2( 1H )-phenanthrone,

3,4,4aer,4-b,5,6,7,8,l0,10aB-Decahydro-4-bfi,8,8- trimethyi-7-oxo-2( 1H )-phenanthrone, and

i0-Acetoxy-8,8-dimethyl3,4,4ao:,4b,8,5,6,7,8,lO,- itia,6decahydro-7oxo-2(li-i)-phenanthrone to give, respectively,

Methyi 3,4,4au,9, l 0, l OawhexahydroJ-methoxy- Ql -phenanthreneacetate [ii; R is (Ii-i R is H, R is CH Methyl 3,4,4ad,4b,8,5,6,7,8,8aoz,9,10,10aB- dodecahydro-7-methyl-7-hydroxy-A -phenanthreneacetateili; R is CH R, R and R" are H, X is H Z is (HO) (CHQCL Methyi 3,4,4ad,4h;8,5,6,7,8,8aa,9,10,1021/3 dodecahydroJcr-methoxyJi -phenanthreneacetate [i; R is CH R", R and R are H, X is H Z is (a "Ci-i U)Ci*i],

32 Methyl 3,4,4a ,4bB,5,6,7,8,8aoi,9,IOJOaBY dodecahydro-7,B-chloro-A -phenanthreneacetate [1; R is CH R", R and R are H, X is H Z is (B.- C1)CH],

Methyl dodecahydrofifi bromo-A -phenanthreneacetate [1, R is CH R", R and R are H, X is H Z is (B.-

Br)CH],

Methyl 4b,8 .8,8-trimethyl-7-oxo-A -phenanth eneacetate [I; R is CH R is H, R is CH R is H, Xis H Z is O=C, A and Methyl 3,4,4aa,4b/3,5,6,7,8,IOJOaB-decahydroJ-oxo- A -phenanthreneacetate [i; R is CH R", R and R" are H, X is (CH;,COO)(H), Z is O=C, A

Example 52 oxo-A -phenantheneacetic acid (isomer A) [i; R, R, R and R" are H, X is H Z is O=,C]

A solution of 6.0 g. (0.021 mole) of ethyl 3,4,4aa,- 4bB,5,6,7,8,8ao1,9,l0,1OaB-dodecahydro-7-oxo- A -phenanthreneacetate (isomer A) (Example 34a) in 200 ml. of 95 percent ethanol was treated with 80 ml. (0.16 mole) of 2N aqueous sodiumhydroxide and the solution was refluxed for minutes under nitrogen, The reaction mixture was cooled, acidified with acetic acid and concentrated under reduced pressure until the ethanol was removed. The product was. ex- 2 tracted from the resulting mixture with ether and then extracted from the ether with 2N aqueous sodium hydroxide. Acidification of this extract with concentrated hydrochloric acid precipitated the product which was collected and recrystallized from ethyl acetate to give w 4.2 g. of 3,4,4aa,4b,8,5,6,7,8,8a(x,9,lO,lOaB-dodecahydro-7-oxo-A -phenanthreneacetic acid (isomer A), m.p. 224227C. (vac) EXAMPLE 53 1 ,2,3,4,4a04,4b,8,5,6,7,8,8aa,9,1O,lOa[3-Tetradecahy+ dro-"IB-hydroxy-Z-pher1anthreneacetic acid (isomer A) [i; R, R, R and R are H, X is H Z is (B-HO)CH, saturated side chain] cording to the procedure of Example 52 and the product was recrystallized once from ethyl acetate ,to give 3.95 g. (97 percent) of l,2,3,4,4aa,4bfi,5,6,7,8- ,8aa,9,1 0,10aB-tetradecahydro-7B-hydroxy-2-phenam threneacetic acid (isomerv A), m.p. 2 l4-216C.'

dro-7B-hydroxy-2-phenanthreneacetic acid (isomer- The mother liquor residues from Example 35 were hydrolyzed according to the procedure of Example 52.

Two recrystallizations of the product from ethyl acetate and one from acetone afforded 3.13 g. of l,2,3,4,4aa,4b/3,5,6,7,8,8aa,9,l0,10aB-tetradecahy- 3,4,4aa,4b;3,5,6,7,8,8aa,9,10,10aB- I 3,4,4aa,4b,5,6,7,8,IOJOaB-decahydro I l0-acetoxy-8,8-dimethylethyl The ester of Example 35 (4.5 g.) was hydrolyzed ac- I EXAMPLE 54 3,4,4aa,9,l O,l0aB-Hexahydro-7-methoxy-A phenanthreneacetic acid [11; R and R are H, R is CH m.p. l74l-75C. (from acetone-hexane).

EXAMPLE 55 3,4,4aa,4b,5,6,7,8,8aa,9,10,10aB-Dodecahydro- 4bB-methy1-A -phenanthreneacetic acid [1; R and R are H, R is CH R is H, X is H Z is CH m.p. 169179C. (from ethyl acetate).

EXAMPLE 56 3 ,4,4aa,4bB,5,6,7,8,8aa,9,10,10aB-Dodecahydro-7- oxo-A -phenanthreneacetic acid (lsomer B) [1; R, R, R and R are H, X is H Z is O=C], m.p. 220-222C. (vac.) (from ethyl acetate).

EXAMPLE 57 3,4,4aa,4bB,5,6,7,8,8aa,9,10,10aB-Dodecahydro- 7a-hydroxy-A -phenanthreneacetic acid [1; R, R, R and R are H, X is H Z is (a-HO)CH], m.p. l73189C. (from ethyl acetate).

EXAMPLE 58 3,4,4aa,4bB,5,6,7,8,8aa,9,10,10aB-Dodecahydro- 7B-hydroxy-A "-phenanthreneacetic acid [1; R,

.R, R and R" are H, X is H Z is (B-HO)CH], m.p.

,205207C. (from ethyl acetate).

EXAMPLE 59 3 ,4 ,4aa,4bB,5 ,6,7,9, 1 0, l 0a/3-Decahydro-7-oxo- A -phenanthreneacetic acid [1; R, R, R and R' are H, X is H Z is O=C, A m.p. l94200C. from ethyl acetate).

I EXAMPLE 60 3 ,4,4aa,4bB,5 ,6,7,8 ,8aa,9,10,10aB-Dodecahydro- 7,7-ethylenedimercapto-A -phenanthreneacetic acid [1; R, R, R and R'are H, X is H Z is .ethylenedithio m.p. l87220C. (from acetone).

EXAMPLE 61 3 ,4,4aa,4b,5 ,6,7,8 ,8aa,9,10,10aB-Dodecahydro- 4b/3-methyl-7-oxo-A -phenanthreneacetic acid [1; R and R are H, R is CH R" is H, X is H Z is O=C], Isomer A, m.p. 18l184C. (from ethyl ace- :tate); and Isomer B, m.p. 219-22lC (from ethyl acetate).

EXAMPLE 62 3,4,4aa,4b,5,6,7,8,8aa,9,10,10aB-Dodecahydro- 4bB-methyl-7B-hydroxy-A a -phenanthreneacetic acid [1; R and R are H, R is CH;,, R" is H, X is H Z is (B-HO)CH], lsomer A, m.p. 223225C.; and Isomer B, m.p. l98200C. (from ether).

EXAMPLE 63 3,4,4aa,4b,5,6,7,8,8aa,9,lO,lOaB-DodecahydrolB,4bB-dimethyl-7B-hydroxy-A -phenanthreneacetic acid [1; R and R are H, R and R are CH X is H Z is (B-HO)CH].

34 EXAMPLE 64 3,4,4a ,4bB,5,6,7,8,8aB,9,lO,lOaB-Dodecahydro-7- oxo-A -phenanthreneacetic acid [1; R, R, R and R are H, X is H Z is O=C, A/B cis], m.p. l85200C. (from ethyl acetate).

EXAMPLE 65 3,4,4aa,4bB,5,6,7,8,8aa,9,10,10aa-Dodecahydro- 7B-hydr0xy-A -phenanthreneacetic acid [1; R,

R, R and R" are H, X is H Z is (B-HO)CH, B/C cis].

EXAMPLE 66 3,4,4aa,4b,5,6,7,9,10,10aB-Decahydro-4bfihydroxy-7-oxo-A -phenanthreneacetic acid [1; R and R are H, R is OH, R is H, X is H Z is O=C, A m.p. 208-232C. (from acetone).

EXAMPLE 67 3,4,4aa,4b/3,5,6,7,8,8aa,9,lO,lOaB-Dodecahydro- 7B-hydroxy-l0-oxo-A -phenanthreneacetic acid [1; R, R, R and R are H, X is 0, Z is (B-HO)CH], Isomer A, m.p. 237238C. (from acetone); and Isomer B, m.p. 282-283C. (vac.) (from acetone).

EXAMPLE 67A 3,4,4aa,4b,5,6,7,8,8aa,9,10,10aB-Dodecahydro-4B- methyl-7B-hydroXy-A -phenanthrenepropionic acid [1; R is H, R and R are CH R" is H, X is H Z is (B-HO)CH].

EXAMPLE 67B 3,4,4aa,4b,5,6,7,8,8aa,9,lO,l0aB-Dodecahydro- 4bB,8,8-trimethyl-7B-hydroxy-N -phenanthreneacetic acid [1; R and R are H, R is CH R is H, X is H Z is (B-HO)CH, 8,8-(C1-1 m.p. l86l90C (from acetone).

According to the procedure of Example 52 the following acids can be prepared by hydrolysis of the corresponding lower-alkyl esters:

3,4,4aa,9,l0,l0aa-Hexahydro-7-methoxy-A phenanthreneacetic acid [11; R and R are H, R" is 3],

3,4,4aa,4bfl,5,6,7,8,8aa,9,lO,lOaB-Dodecahydro-7- methyl-7-hydroxy-A -phenanthreneacetic acid [1; R, R, R and R are H, X is H Z is (HO)(CH )C],

3,4,4aa,4b[3,5,6,7,8,8aa,9,l0,IOaB-Dodecahydro- 7a-methoxy-A -phenanthreneacetic acid [1; R, R, R and R are H, X is H Z is (B-CH -,O)CH],

3,4,4aa,4bB,5,6,7,8,8a0z,9,l0,10aB-Dodecahydro- 7B-fluoro-A a -phenanthreneacetic acid [1; R, R, R and R are H, X'is H Z is (B-F)CH],

3,4,4aa,4b,6,5,6,7,8,8aa,9,10,10aB-D0decahydro- 7B-chloro-A -phenanthreneacetic acid [1; R, R, R and R are H, X is H Z is (B-CUCH],

3,4,4aa,4bfl,5,6,7,8,8aa,9,10,10a/3-Dodecahydro- 7B-bromo-A a -phenanthreneacetic acid [1; R, R, R and R are H, X is H Z is (B-Br)CH],

3,4,4aa,4b,5,6,7,8,l0,IOaB-Decahydro-4bfi,8,8- trimethyl-7-oxo-A -phenanthreneacetic acid [1; R and R are H, R is CH R is H, X is H Z is O=C, A8113],

3,4,4aa,4bB,5,6,7,8,l0,lOaB-Decahydro-7-oxo-l0- hydroxy-A -phenanthreneacetic acid [1; R, R, R and R are H, X is (H)(OH), Z is O=C, A and EXAMPLE 68 7,7Ethylenedimercapto-3,4,4a0z,4b,5,6,7,8,8aa,9,10,- llafi-dodecahydro-4bB-rnethyl-A -phenanthreneacetic acid [1; R and R are H, R is CH R" is H, X is H Z is (ethylenedithio)C] A solution of 1.64 g. (5.9 mmoles) 3,4,4ac,4b,5,6,7,8,8a0z,9,l0,10afl-dodecahydro4bfimethyl-7-oxo-A -phenanthreneacetic acid (EX- ample 61) in 15 ml. of acetic acid was treated with 2.0 m1. of ethanedithiol followed by 2.0 ml. of boron trifluoride etherate. No heat was evolved but a crystalline precipitate formed immediately. After minutes the mixture was diluted with mi. of water and filtered. The filter cake was washed well with water, air-dried and recrystallized from 100 ml. of acetic acid to give 1.56 g. of 7,7-e'thylenedirnercapto-3,4,4aa,4b,5,6,7,8- ,8a0z,9,10, iOaB-dodecahydro-Abflmethyl-A a phenanthreneacetic acid, m.p. 254-259C.

By replacing the ethanedithiol in the foregoing prep aration by a molar equivalent amount of ethylene gly col, 1,3-propylene glycol, or l,3-propanedithiol, there can be obtained, respectively, 7,7-ethylenedioxy- 3 ,4,4ao,4b,5 ,6,7,8,8aa,9,10,10aB-dodecahydro-4b18- methyl-A -phenanthreneacetic acid [1; R and R" are H, R is CH R"is H, X is H Z is (ethylenedioxy)C], 7,7-(1,3-propylenedioxy)-3,4,4a0r,4h,5,6,7,8- ,8aa,9,10,i09.,6-dodecahydro-4bB-methyl-A phenanthreneacetic acid H; R and R are l i, R is CH R is 11", X is i-l Z is (l,3-propylenedioxy)C], or 7,7- (1,B-propylenedirnercapto)-3,4,4a0:,4b,- 5,6,'7,8,8a0z,9,10,10aB-dodecahydro-4bfi-rnethyl- M -phenanthreneacetic acid [1; R and R" are H, R is Ii-i R" is H, X is H Z is (1,3propylenedithio)].

Similarly, 3,4,4a01',4b,5,6,7,8,8aa,9.10,10afidodecahydro-4bB-methyl-7-oxo-A -phenanthreneacetic acid was caused to react with benzylmen captan to give 7,7-dibenzylmercapto- 3,4,4aor,4b,5,6,7,8,8aa:,9,10,l0aB-dodecahydro-4bB- methyl d -phenanthreneacetic acid [1; :R and R are H, R is CH R" is H, X is l-l Z is (C H Cl-l S) C], mp. 187i94C.

EXAMPLE 69 7,7-(Ethylenedisulfonyl)-3,4,4a01,4b,8,5,6,7,8- ,82.C6,9,iO,IOZB-dOdeCQhYdI'O-A -phenanthreneacetic acid [1; R, R, R and R are H, X is H Z is (ethylenedisulfonyl)C] A solution of 1.2 g. (3.5 mmoles) of 7,7- (ethylenedirnercapto)-3,4,4a0z,4b,8,5,6,7,8,8a0z,9,10,- llflafi-dodecahydro-A -phenanthreneacetic acid (Example 60) in 150 ml. of ether was treated wtih 3.08 g. (17mrnoles) of monoperphthalic acid in 21 ml. of ether. The solution was kept overnight at room temperature, 150 ml. of tetrahydrofuran was added and the solution was left for three more days. Ether (500 ml.) was added and the solution was washed with saturated sodium sulfite solution and brine and then dried over sodium sulfate. The solution was concentrated to give a crystalline residue which was triturated with about ml. of chloroform and collected on a filter; 1.13 g., m.p. 266267C. (dec.). A second crop of 0.19 g., mp.

3(6) 267*270C., was obtained by concentration of the chloroform washings (92 percent yield). Recrystalliza- 0 tion from acetone gave 7,7-(ethyienedisulfonyl)- 3,4,4216t,4i3,5,6,7,8,82l0,9,i0,iOaB-fiOCifiCahYCilO M 85 -phenanthreneacetic acid, mp. 270,271C.

Similarly, 7,7-dibenzyimercapto-3,4-,4-a0z,4b,5,6,7,8-

EXAMPLE 3,4-,4ac ,4b,5,6,7,8,8ao,9,10,10aB-Uodecahydr0-4bfimethyi-7B-pyrrolidino43 a -phenanthreneacetic acid [1;R and PC are H, R is (11-1 is H, X is H Z ample 61), ml. of benzene and 8 ml. (96 mmoles) of pyrrolidine was heated under reflux for four and onehalf hours with a water separator attached to the system. This solution was concentrated to aresidue by warming under reduced pressure and theresidue was treated with 50 ml. of dry benzene and 3.5 ml. (93

rnmoles) of formic acid. The mixture was heated under reflux for 30 minutes, cooled and treated with 1.5 mi. of formic acid. Water (60 ml.) and ether (100 ml.) were added and the layers were separated. The ether layer was extracted once with 2N hydrochloric acid and discarded. Addition of the acidic extract to the aqueous portion of the reaction mixture caused precipitation of the hydrochloride salt of the product. Concentrated hydrochloric acid (3nd.) and 10 ml. of brine were added and the precipitate was collectedlt was washed well with acetonitrile and then ether to give threneacetic acid which was suitable for conversion to a basic ester.

By replacing the pyrrolidine in the foregoing prepara tion by a molar equivalent amount of diethylamirie or piperidine there can be obtained, respectively, 3,4,4a0i,4b,5,6,7,8,8aoz,9,l0,10aB-dodecahydro-4bfimethyl-7B-diethylamindd -phenanthreneacetic acid [1; R and R are H, Ris (Ill-l R is H, X is H Z is (C H N]Cll-i, or 3,4,4ad,4b,5,6,7,8,8a0r,9,10,10aB- dQdecahydrorbB-methyl7B-piperidino-A phenanthreneacetic acid [i; R and R are H, R is CH R is H, X is H Z is (CH NCH].

EXAMPLE 71 8,8-Dimethyl-7, l 0-dioxo-3,4,4ao. ,4bB,5,6,7,8, 10,- I

R, R", R and R are lhl, X is 0, Z is'O=-C, 8,8-(CH Ai can be prepared from the corresponding methyl ester (Example 51) by methods which do not open Ring A, for example, by heating the methyl ester with anhydrous lithium iodide in the presence of a suitable solvent such as collidine.

Conventional aqueous alkaline hydrolysis Oflth methyl ester leads to a compound where Ring A has been opened as follows:

Methyl 8,8-dimethyl-3,4,4a0z,4b,8,5,6,7,8,10,10aB- decahydro-7,lO-dioxo-A -phenanthreneacetate (11.2 rnrnoles) (lsomer A) (Example 51) (810 mg., 2.5 millimoles) in 20 ml. of ethanol and 8 ml. of 2N sodium hydroxide was boiled under reflux in a nitrogen atmosphere for one hour. The reaction mixture was added to ice-water and was made acidic with dilute hydrochloric acid. Sodium chloride was added to the reaction mixture which was then extracted with ether. The ether was dried (Na SO and evaporated to afford 810 mg. of residue. The residue was chromatographed on 100 g. of silica gel. Elution with acetic acid-ether-pentane (3:50:47) afforded 230 mg. of 5a-(2-carboxyethyl)-3,4,4aa,5,8- ,SaB-hexahydro-6-isopropyl-8-oxo-A -naphthaleneacetic acid (lsomer A), rn.p.'l86l90C. Recrystallization from ether gave a sample that melted at l94-l95C.

Similarly, methyl 8,8-dimethyl- 3 ,4,4aa,4bB,5,6,7,8,10,10aB-decahydro-7,lO-dioxo- A -phenanthreneacetate (lsomer B) (Example 51) was hydrolyzed to a-(2-carboxyethyl)- 3,4,4aoz,5,8,8aB-hexahydro-6-isopropyl'8-oxo- A -naphthalenacetic acid (lsomer B), 2l32l5C. (from ether).

5a-(2-Carboxyethyl)-3,4,4aa,5,8,8aB-hexahydro-6- isopropyl-8-oxo-A -naphthaleneacetic acid (Isomers A and B) were found to have antibacterial activity when tested in vitro against conventional test organisms such as Staph. aureus and E. typhi.

EXAMPLE 72 Z-Dimethylaminoethyl 3,4,4aa,4bB, 5,6,7,8,8aa,9,10,10aB-dodecahydro-7- oxo-A -phenanthreneacetate (Isomer A) [1; R is CH CH N(CH R, R and R are H, X is H 2 is A solution of 4.47 g. (0.017 mole) of the 3,4,4aa,4bB,5,6,7,8,8aa,9,l0,lOaB-dodecahydro-7- oxo-A -phenanthreneacetic acid (Isomer A) (Example 52) in 100 ml. of tetrahydrofuran was treated with 0.92 g. (0.17 mole) of sodium methoxide and 1 ml. of water. The solvent was then removed by warming under reduced pressure, 20 ml. of absolute ethanol was added and evaporated in the same manner and, finally, two 20-ml. portions of dry benzene'were added and evaporated. The resulting dry sodium salt was suspended in l50 ml. of dry benzene, 3.46 g. (0.044 mole) of pyridine was added, the mixture was immersed in an ice bath and 40 ml. of oxalyl chloride was added in a fast stream of drops with stirring. The mixture was removed from the ice bath, stirred for 10 minutes and thenconcentrated as rapidly as possible under reduced pressure using a water bath at 45C. Application of heat was stopped as the last of the solvent evaporated, and 150 ml. of benzene was added followed by 40 ml. of Z-dimethylaminoethanol in a rapid stream of drops with stirring and cooling. When addition was complete, the mixture was heated on the steam bath for 5 minutes, cooled and diluted with 1 liter of ether and 600 ml. of saturated aqueous sodium carbonate. The layers were separated and the water layer was washed with ether and discarded. The combined ether layers were extracted with two IOO-ml. portions and one SO-ml. portion of 2N hydrochloric acid 38 a 96 percent trans-4 percent cis mixture of isomers to gether with 12 percent of impurity.

The product was purified by partition chromatogra phy. The solvent system employed was a l2:l:2:O.2 mixture of hexane, ethylene dichloride, methanol and water. lnfusorial earth (300 g.) was wet with 225 ml. of the polar phase containing mg. of bromcresol purple, the color of the mixture was adjusted to a pale creamy yellow (faintly acid) by gaseous hydrogen chloride and the solid was packed into a column 9 cm. in diameter. The sample was dispersed on 10 g. of infusorial earth and placed on the top of the column. Elution of the column with the non-polar phase of the solvent mixture developed the column; the position of all basic material was clearly revealed by blue bands. The product was recovered either by elution or slicing of the column, depending on the separation of the bands. In the present case the product was eluted to yield 4.39 g. of the basic ester mixture which was free of significant impurities.

The 4.39 g. of oil was dissolved in 200 ml. of ether and treated with 1.1 1 ml. of concentrated hydrochloric acid in 20 ml. of absolute alcohol. The precipitated hydrochloride salt was collected and recrystallized twice 'from acetone to give 3.56 g. of Z-dimethylaminoethyl 3,4,4a ,4bB,5,6,7,8,8aa,9,l0,10aB-dodecahydro-7- oxo-A -phenanthreneacetate (isomer A) in the form of its hydrochloride salt, colorless plates, m.p. l80.0l82.0C.

Similarly there was prepared Z-dimethylaminoethyl 3,4,4aa,4bB,5,6,7,8,8aa,9,l0,10aB-dodecahydro-7- oxo-A -phenanthreneacetate isomer B, hydrochloride salt, m.p. l74.5l75.5C. (from acetone) containing 541 mole of water of crystallization.

By replacing the Z-dimethylaminoethanol in the foregoing preparation by a molar equivalent amount of 2- (l-piperidyDethanol, 2--( l-pyrrolidyl)ethanol, 2-( 4- morpholinyl)ethanol, 2-(4-methyl-l-piperidyl)ethanol, or 2-(4-methyl-l-piperazinyl)ethanol, there can be obtained, respectively, 2-( l-piperidyl )ethyl 3,4,4aa,4bB,5,6,7,8,8aot,9,l0,l0aB-dodecahydro-7- oxo-A -phenanthreneacetate [1; R is CH CH N(CH R, R and R and H, X is H Z is O=C], 2-(l-pyrrolidyl)ethyl 3,4,4aoz,4bfi,5,6,7,8- ,8aoz,9,l0,1OaB-dodecahydro-"l-oxo-A a phenanthreneacetate [l; R is CH CH N(CH R, R and R are H, X is H Z is O=C], 2-(4-morpholinyl- )ethyl 3,4.4aa,4b,8,5,6,7,8,8ao,9,lO,10a,8-dodecahydro-7-oxo-A -phenanthreneacetate [l; R is CH CH N(CH O, R, R and R are H, X is H Z is O=C], 2-(4-methyl-l-piperidyl)ethyl 3,4,4aa,4bB,5,6,7,8,8aa,9,l0,10aB-dodecahydro-7- oxo-A -phenanthreneacetate [1; R is CH CH N(CH CHCH R, R and R" are H, X is H Z is O=C], or 2-( 4-methyl-l-piperazinyl)ethyl 3,4,4aa,4bB ,5,6,7,8,8a0z,9,10,10aB-dodecahydro-7- oxo-A -phenanthreneacetate (l; R is CH CH N(CH NCH R, R and R are H, X is H Z is O=C].

By the foregoing esterification procedure the following examples of basic esters (R is amino-lower-alkyl) were prepared from the corresponding acids (R is H):

EXAMPLE 73 Z-Diethylaminoethyl 3,4,4aa,9,l0,lOaB-hexahydro- 7-methoxy-A -phenanthreneacetate [l]; R is CH CH N(CH R is H, R is CH hydrochloride salt, m.p. l79183C. (from acetone). 

1. 1,2,3,4,4A,4B,5,6,7,8,8A,9,10,10A-TETRADECAHYDROPHENANTHRENE BEARING IN BOTH THE 2- AND 7-POSITONS A HYDROXY GROUP, AND WHEREIN THE 4B-POSITION IS UNSUBSTTITUTED OR BEARS A LOWER-ALKYL GROUP.
 2. 1,2,3,4,4a,4b,5,6,7,8,8a,9,10,10a-Tetradecahydrophenanthrene-2,7-diol, according to claim 1, wherein there are hydroxy groups in the 2- and 7-positions and the 4b-position is unsubstituted.
 3. 1,2,3,4,4a,4b,5,6,7,8,8a,9,10,10a-Tetradecahydro-4b-Methylphenanthrene-2,7 -diol, according to claim 1, wherein there are hydroxy groups in the 2- and 7-positions and a methyl group in the 4b-position. 